INSTI Training Manual

TRAINING PACK
HIV-1 / HIV-2 Antibody Test
DISTRIBUTED BY
ALSO NOW AVAILABLE INSTI ® SELF TEST

TRAINING PACK
BLOOD
COLLECTION
KIT
MULTIPLEX HIV-1/HIV-2
Syphilis Ab Test Kit
Product code: 8400
NHS Supply Chain Code: HHH2286
HIV-1/HIV-2
Antibody Test Kit
Product code: 8300
NHS Supply Chain Code: HHH208
www.hivhometest.com
www.biolytical.com

Special INSTI ® Features
• Built-in true human IgG Control
• Use 50μL of EDTA-whole blood, fingerstick blood, serum or
EDTA-plasma
• Rapid results at a glance (as little as 60 seconds)
• Proven early antibody detection
• Large scale Canadian clinical trial data shows a >99.5%
combined accuracy
• Easy to understand instructions
• No timing required
• No refrigeration required
Commitment to Quality
To ensure the integrity and quality of every test, INSTI ® is
manufactured in a state of the art facility with a fully implemented
Certified Quality Management System (ISO 13485).
Our staff of professionals in chemistry, molecular biology,
biochemistry, engineering and highly skilled technicians adhere
to strict process controls. Our facility is compliant with all
regulations for the manufacture of medical devices.
All raw materials and processed kit components are thoroughly
inspected and tested to ensure they meet strict quality guidelines.
Checks at each step of manufacturing ensure continued quality
control. Rigorous in-process testing using EDTA-whole blood,
plasma and serum is performed during each production run.
All INSTI ® solutions are proprietary formulations allowing us to
control quality in our own laboratories. Our research and
development team continues to develop new diagnostic tests.
ADVANCED
DIAGNOSTIC
TECHNOLOGY
FAST
• Result in 60 seconds
SIMPLE
• All reagents in ready-to-use bottles
• No measuring or drop counting
• No contamination
• No timing required
• No additional materials required
CONVENIENT
• Utilises EDTA-whole blood, fingerstick
blood, serum or EDTA-plasma
ACCURATE
• Excellent correlation with approved
laboratory methods for HIV antibody
detection
SENSITIVE & SPECIFIC
• Recombinant HIV-1 and HIV-2 proteins
offer a >99.5% combined accuracy

Table of Contents
Interpretation of Results/Images 3
Guidelines & Interpretation 4
Guidance for Fingerstick Sample 5
Lateral View of Membrane Unit 6
INSTI ® Do’s and Don’ts & FAQ’s 7
FAQ’s & Summary and Intended Use 8
Troubleshooting 9
Quick Reference 10
True Human IgG Control Spot 11
Instructions for Use 12-15
Test Controls IFU 16
Bibliography 17-18
Clinician’s Briefing Notes 19
Mutliplex Joint Test
Four Simple Steps 20
Interpretation of Results 21
Instructions for Use 22-26
Syphilis Test Control IFU 27-28
Self Test
Introduction 28
Self Test IFU 29
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Interpretation of Results
HIV SPOT INTERPRETATION
Control
Always read
results with
INSTI ® logo
facing you
HIV
Strong Reactive
Weak Reactive
Indeterminate
Negative
SYPHILIS SPOT INTERPRETATION
Control
Always read
results with
INSTI ® logo
facing you
Syphilis
Strong Reactive
Weak Reactive
Indeterminate
Negative
HIV & SYPHILIS SPOT INTERPRETATION
Control
Always read
results with
INSTI ® logo
facing you
Syphilis
HIV
Strong Reactive
Weak Reactive
Syphilis detection is only available on the INSTI ® Multiplex HIV-1/2 Syphils Ab Test.
3

Guidelines & Interpretation
Non-homogeneous appearance of the control dot
Colour intensity may be variable between the ring and the dot.
Please be advised that the appearance of the control dot on the blotted membrane units may be nonhomogeneous
in appearance, with patterns of lighter and darker colour intensities appearing within the intact
dot area as illustrated above. Control dots with this non-homogeneous appearance are valid and should be
interpreted as per the INSTI ® Instructions for Use (package insert).
bioLytical has conducted extensive testing of INSTI ® membrane units and concludes that the INSTI ® membrane
units displaying these non-homogeneous control dots continue to meet all Quality Control and performance
specifications including sensitivity and specificity.
bioLytical has filed vigilance reports on this observation with the appropriate regulatory and competent
authorities.
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Guidelines for obtaining fingerstick sample for an INSTI ®
test
1 Patient to wash hands thoroughly in warm soapy water.
2 With gloves on, milk fingers towards the tip while observing which
finger is optimal - look for the one that has the deepest colour for the
ultimate blood flow.
3 Holding the finger correctly as you use the lancet.
4 Angle the capillary tube horizontally. When collecting blood sample
hold the capillary tube at an angle as shown in the picture. Capillary
action will be assisted by gravity.
5

Lateral view of membrane
Lateral view of Absorbent Pad
During INSTI ® Procedure
1
Sample diluent absorbs
through membrane
into absorbent pad
2
Colour developer
blends with sample in
absorbent pad
3
Clarifying solution
blends with the other
solutions in the pad
Over time, the solutions in the absorbent pad spread uniformly, and
can stain the membrane from below, occasionally resulting in a faint
shadow at the test spot.
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Do’s and Dont’s & Frequently Asked Questions
Do’s
3 Read the entire package insert and become familiar with its contents prior to beginning the test.
3 Ensure all test components and any stored specimens are at room temperature.
3 Warm the fingertip before using the lancet, to ease blood flow.
3 Allow the alcohol from the swabbed fingertip to dry completely before applying the lancet.
3 After addition of blood sample to bottle #1, shake well and immediately pour into the membrane unit.
3 Shake bottle #2 to re-suspend the solution before pouring into the membrane unit.
3 Use local universal precaution guidelines for infectious materials in handling and disposing of
specimens and used testing materials.
Don’ts
7 Do not mix reagents from different lots.
7 Do not use reagents or kits beyond the stated expiration date on the outer packaging.
7 Do not use the test kit if there is any sign of damage to or previous opening of the sealed membrane
pouch, or leakage in any of the reagent bottles.
7 Do not conduct repeated sampling of fingerstick blood with the disposable transfer pipette.
50µL should be collected in one continuous flow.
7 Do not dilute the specimens prior to adding to bottle #1 of the INSTI ® test.
7 Do not add the bottle #1 contents to the membrane unit without mixing well just before addition.
7 Do not add specimen directly to INSTI ® membrane unit.
7 Do not introduce delays between any of the INSTI ® procedural steps.
7 Do not interpret the results if more than 5 minutes have elapsed after adding the Clarifying Solution
(bottle #3) to the INSTI ® membrane unit.
What is the accuracy of the INSTI ® kit?
Proven accuracy of >99.5%
Does INSTI ® differentiate HIV-1 from HIV-2 antibodies?
No. INSTI ® contains antigens from HIV-1 and HIV-2 in the membrane unit and can detect antibodies to HIV-1
and/or HIV-2 in patients’ specimens, but it will not differentiate the two.
Is a reactive INSTI ® test conclusive of HIV diagnosis?
No. INSTI ® is considered as a first-line test only, and all patients with reactive or indeterminate INSTI ® results
should be re-tested with an HIV confirmatory assay before final HIV antibody status can be determined. As in
all assays for HIV antibody, false positives can occur infrequently with INSTI ® .
Are INSTI ® results as accurate as laboratory-based HIV testing procedures?
Yes. INSTI ® is a registered and approved medical device and has undergone extensive clinical trials that have
demonstrated its accuracy in direct comparison to laboratory-based methods. Much of this data is published
in the INSTI ® package insert.
Does INSTI ® detect HIV antibodies early in the course of infection?
Yes. In commercial HIV seroconversion panels and ‘real life’ documented seroconversion cases, INSTI ® is
equivalent to the most sensitive laboratory-based assays in early antibody detection. However, patients who
are in the pre-antibody “window period” of HIV infection may not be reactive in the INSTI ® test.
7

Frequently Asked Questions & Summary and Intended Use
What kind of antigen is used?
The INSTI ® test utilises recombinant transmembrane proteins from HIV-1 and HIV-2. Use of these proteins
overcome sensitivity and specificity problems associated with a test based on viral lysates or a combination of
core antigen and other viral proteins.
Does the INSTI ® kit have a built-in sample addition control?
Yes, the procedural control for the INSTI ® kit consists of a protein capable of capturing the IgG normally present
in human blood components. IgG is present in blood components in both HIV negative and HIV positive human
specimens. Captured IgG reacts with a chromatic agent to produce a visual sign (dot) on the control membrane.
Does the INSTI ® kit only react with whole blood samples?
No, you can use the INSTI ® kit with EDTA-whole blood, EDTA-plasma, fingerstick, serum or plasma
samples.
How long does it take to perform a single test?
INSTI ® preparation is minimal and results show up in as little as 60 seconds. Results are interpreted immediately
after pouring the clarifying solution (bottle #3) into the membrane unit.
What is the stability of the test results in the test membrane unit?
The test results will remain visible in the INSTI ® membrane unit for an indefinite period after performing the
test. Read within the first 5 minutes after the test. The results may fade over time.
What kind of storage conditions are needed?
INSTI ® tests should be stored at:
1. INSTI ® HIV-1/HIV-2 Antibody Test kit: store at 2-30°C
2. INSTI ® HIV-1 and HIV-2 Controls: Store frozen at -20°C or refrigerate at 2°C to 8°C for up to one year. Once
thawed store at (2- 8°C). They remain stable until expiry (up to 1 year). Do not re-freeze once the bottles have
been opened.
3. INSTI ® Multiplex Test kits: store at 15-30°C
4. INSTI ® Syphilis Controls: Store at 2 - 8°C for up to 28 days or at ≤-20°C for up to 1 year. Once the controls are
thawed following storage at ≤-20°C, they remain stable for 28 days at 2 - 8°C. Do not re-freeze once the
bottles have been opened.
5. INSTI ® HIV Self Test: store at 2-30°C
Note - the storage conditions for INSTI ® HIV-1/HIV-2 Antibody Test kits and INSTI ® Multiplex Test kits are
different. Also the storage conditions for HIV and Syphilis controls are different.
What is the shelf life?
INSTI ® tests presently have a shelf life of:
1. INSTI ® HIV-1/HIV-2 Antibody Test kit: Shelf-life of 15 months from date of manufacture
2. INSTI ® HIV-1 and HIV-2 Controls: Shelf-life of 12 months from date of manufacture
3. INSTI ® Multiplex Test kit: Shelf-life of 12 months from date of manufacture
4. INSTI ® Syphilis Controls: Shelf-life of 12 months from date of manufacture
5. INSTI ® HIV Self Test: Shelf-life of 15 months from date of manufacture
What level of quality assurance do you practice in your manufacturing facility?
bioLytical ® Laboratories is an ISO 13485 certified manufacturing facility that has been inspected and approved
by the FDA, European Notified Body and Health Canada for the manufacturing of medical devices. Each INSTI ®
test kit undergoes multiple quality assurance steps and procedures prior to being released for sale.
What are the special benefits of using INSTI ® ?
• Results in 60 seconds
• Proven accuracy of >99.5%
• Proven early detection
• Built in sample addition control
• Easy to use and understand
Where can I learn more about bioLytical ® Laboratories and INSTI ® ?
For enquires or to learn more, please visit our website at: www.biolytical.com, insti.com or www.pasante.com
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Troubleshooting
Not for donor screening
The INSTI ® HIV-1/HIV-2 Antibody Test is a single use, rapid, flow-through in vitro qualitative immunoassay for
the detection of antibodies to Human Immunodeficiency Virus Type 1 and Type 2 in human EDTA-whole blood,
fingerstick blood, serum or EDTA-plasma.
The test is intended for use by trained personnel in medical facilities, clinical laboratories, emergency care
situations, and physicians’ offices as a diagnostic test capable of providing results in less than one minute.
Although suitable for near-patient or point-of-care (POC) testing, the INSTI ® HIV Antibody Test is not suitable
for home testing at this time.
All required pre and post-test counselling guidelines must be followed in each setting in which the INSTI ® HIV
Antibody Test is used. The assay is packaged as a kit containing an INSTI ® Membrane Unit, Sample Diluent,
Colour Developer, and Clarifying Solution with or without support materials (lancet, pipette and alcohol swab).
Invalid Results:
No dots: Usually an indication that substantially less than 50μL of specimen (blood, serum or plasma) was used.
Could also be an indication that the INSTI ® solutions were added in the wrong sequence. Testing should be
repeated with a fresh kit.
Solid Colour: A uniform blue tint across the entire membrane which obscures the control and test dots is an
indication that the reagents were not used in the correct sequence. Testing should be repeated.
Specks on the membrane: This could be caused by slight aggregation of the contents of bottle #2. If the control
dot is clearly visible, test results are valid even with specks on the membrane. Specks can also appear on the
membrane if the kit was used past the expiry date, in which case the results are not considered valid.
Test dot only: This might occur very rarely and may be an indication that the test procedure has not been
followed properly or there was a manufacturing problem. Testing should be repeated, on a fresh kit.
Additional factors which may cause Invalid results:
• The membrane unit has not been used immediately after opening.
• Reagents from different kit lots are combined to perform the INSTI ® test.
• Using reagents in the wrong order.
• Some reagents are spilled while pouring into the membrane unit.
• The INSTI ® kits were stored at temperatures outside the approved storage temperature range.
• The membrane has been contaminated causing blockage of flow.
Indeterminate Results:
In some instances, test dots might appear as very faint shadows or blue rings. These are to be interpreted as
Indeterminate results, and testing should be repeated. Consult the INSTI ® package insert section on
Interpretation of results for follow-up action.
Indeterminate or Invalid results may occur if:
1. More or less than 50μL of blood, serum or plasma is added to the test.
2. Flow times for the reagents are slow or greater than 2 minutes in total.
3. There are delays introduced in the procedural sequence once the sample is added to bottle #1.
Or delays between pours.
4. The contents of bottle #1 and bottle #2 are not mixed immediately before pouring into the membrane unit.
5. INSTI ® kits are used beyond the expiry date.
9

HIV
Quick Reference Guide
(1) (2) (3) (4)
» add 50µL
(blood, serum, plasma)
» shake well and pour
» allow to flow through
» shake well and pour
» allow to flow through
» allow to flow through
» read immediately
(within 5 minutes)
Procedure Steps (1-4) are to be carried out in sequence with no delays. If there is any delay between
Procedure Steps (1) and (2):
a) Step 2 must be performed within 5 minutes after Step 1.
b) Re-mix Sample Diluent (bottle #1) by inverting several times before pouring into the Membrane Unit.
c) Immediately proceed to Steps 3 and 4.
Do not conduct repeated sampling involving multiple transfers of small amounts of blood from fingertip to
Sample Diluent Bottle #1 as this may result in improper blood volumes.
To facilitate fingerstick blood collection, the fingertip at the collection point should be warm.
Ensure that the fingertip is completely dry after the use of the Alcohol Swab, before applying the Lancet.
Test
Control
Test
Control
Control
Test
Invalid
Indeterminate
Non-Reactive Reactive
A Reactive Test result should be readily visible to the naked eye. Repeat the test if the results are unclear.
Following a reactive or indeterminate INSTI ® test result, a venous blood sample must be drawn in a
‘lavender-top’ EDTA collection tube (for whole blood or plasma) or red-top tube (for serum), and forwarded
to laboratory of HIV confirmatory testing.
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True Human IgG Control Spot
Built-In Sample Addition Control:
The INSTI ® HIV-1/HIV-2 Antibody Test kit has a built-in sample addition control capture that demonstrates assay
validity and adequate sample addition. A blue colour in the control spot indicates that the proper specimen
was added and that the assay procedure was performed correctly. The control spot will be visible on all valid
INSTI ® tests.
With INSTI ® , if the control dot appears, this indicates the following:
• The test has been performed correctly.
• The flow through mechanism of INSTI ® has worked properly.
• The correct amount and type of specimen has been added to the membrane unit.
• In the absence of a visible test dot, this is a valid non-reactive result.
• Any test dot that is readily visible is a valid reactive result.
External Controls:
Only INSTI ® controls should be used with INSTI test kits. It is advisable that external control material, consisting
of at least one HIV negative and one HIV positive serum or plasma specimens, be tested with INSTI ® :
• At regular intervals or with every shipment of INSTI ® .
• For new INSTI ® user verification.
• When switching to new lot number of INSTI ® test kits.
• When the storage area falls outside 2-30˚C.
It is the responsibility of each user of the INSTI ® HIV-1/HIV-2 Antibody Test kit to establish an adequate quality
assurance program to ensure proper performance under their specific locations and conditions of use.
11

HIV
Instructions for Use
Y
HIV-1/HIV-2 Antibody Test
Single-use rapid assay for the detection of antibodies to
Human Immunodeficiency Virus Type 1 (HIV-1) and Type 2 (HIV-2)
90-1015 - One INSTI ® HIV-1/HIV-2 Antibody Test with support materials (for POC use)
Store at 2°C to 30°C
K
Caution
Harmful if swallowed g
Lot number
Sterilization using irradiation
V
In Vitro diagnostic medical device
h
Catalogue Number
i Consult instructions for use M Manufacturer
D Do not reuse C CE Mark
H
Use by
Store at 2°C – 30°C. For in vitro diagnostic use only. V
i It is recommended that the entire Instructions for Use be read prior to beginning the test
procedure. Although the assay is designed to be simple to use, conformance with the test procedure is
necessary to ensure accurate results.
INTENDED USE - Not for donor screening
The INSTI HIV-1/HIV-2 Antibody Test is a single use, rapid, flow-through in vitro qualitative
immunoassay for the detection of antibodies to Human Immunodeficiency Virus Type 1 and Type 2 in
human EDTA whole blood, fingerstick blood, serum or EDTA-plasma. The test is intended for use by
trained personnel in medical facilities, clinical laboratories, emergency care situations, and physicians’
offices as a diagnostic test capable of providing results in less than one minute. Although suitable for
near-patient or point-of-care (POC) testing, the INSTI HIV-1/HIV-2 Antibody Test is not intended for
home testing. All required pre, and post-test counselling guidelines must be followed in each setting in
which the INSTI HIV-1/HIV-2 Antibody Test is used. The assay is packaged as a kit containing INSTI
Membrane Unit, Sample Diluent (Solution 1), Colour Developer (Solution 2), and Clarifying Solution
(Solution 3) with support materials (lancet, pipette and alcohol swab).
SUMMARY
Acquired Immunodeficiency Syndrome (AIDS) is caused by at least two retroviruses, HIV-1 and HIV-2.
HIV-1 and HIV-2 are similar in genomic structure, morphology and ability to cause AIDS. 1 HIV is
transmitted mainly by sexual contact, exposure to blood or blood products, or from an infected mother
to her foetus. People with increased risk of HIV infection include haemophiliacs, intravenous drug-users
and men having sex with men (MSM). HIV has been isolated from patients with AIDS, AIDS-related
complex (ARC), and from persons at high risk of contracting AIDS. 2-5 Antibodies specific for HIV
envelope proteins are prevalent in sera from persons at high risk of contracting AIDS as well as in people
with AIDS, or ARC. 5-7 The presence of antibodies to HIV indicates previous exposure to the virus, but
does not necessarily constitute a diagnosis of AIDS. The prevalence of antibodies to HIV in people not
known to be at risk of acquiring HIV infection is unknown, but significantly less. 5 Absence of antibodies
to HIV does not indicate that an individual is free of HIV-1 or HIV-2; HIV has been isolated from
seronegative individuals prior to seroconversion. Test specificity and sensitivity depend, amongst
other factors, on: a) the selection of HIV antigens used for antibody detection, b) the classes of
antibodies recognized by the detection conjugate, and c) complexity of the protocol used to perform the
test. 8 Non-specific reactions may be observed in some specimens. A reactive INSTI test result should
be considered a preliminary result, with appropriate counselling provided in POC settings. Following a
reactive rapid test result, a venous blood sample must be drawn in an EDTA collection tube (for whole
blood or plasma) or a tube with no anticoagulant (for serum), and forwarded to a laboratory for HIV
confirmatory testing.
PRINCIPLES OF THE TEST
C
0543
The INSTI HIV-1/HIV-2 Antibody Test is a manual, visually read, flow through immunoassay for the
qualitative detection of HIV-1/HIV-2 antibodies in human blood, serum or plasma. The test consists of a
synthetic filtration membrane positioned atop an absorbent material within a plastic cartridge, referred
to as the INSTI Membrane Unit. The membrane has been specifically treated with HIV-1 and HIV-2
recombinant proteins, which react with HIV-1/HIV-2 antibodies in the specimen to produce a distinct
visual signal on the membrane. The membrane also includes a procedural control. The procedural
control consists of a protein-A treated spot capable of capturing IgG antibodies normally present in blood
and blood components. IgG antibodies react with a proprietary chromatic agent to produce a visual
signal on the membrane. Since IgG antibodies are present in blood from normal or HIV positive human
specimens, the control dot provides a visual signal when the test is run, indicating that the test was
performed correctly. If the control dot does not appear, the test is considered invalid. In the case of the
test spot, recombinant HIV-1 and HIV-2 proteins, embedded in the membrane, capture HIV specific
antibodies, if present in the specimen. Antibodies captured in the test spot react with a proprietary
chromatic agent to produce a visible signal on the membrane. The membrane unit is designed to filter,
absorb, and retain the test specimen and all the test reagents in such a manner as to limit leakage and
exposure of personnel to potentially infectious materials.
Reagents required to conduct a test include Sample Diluent (Solution 1), Colour Developer (Solution 2)
and Clarifying Solution (Solution 3). The test is performed by adding the blood, serum, or plasma
specimen to the vial of Sample Diluent, which lyses the red blood cells. This specimen/diluent solution
is then poured onto the well of the Membrane Unit. HIV-1/HIV-2 antibodies, if present in the specimen,
are captured by proteins on the filtration membrane. Colour Developer is then added to the Membrane
Unit. The Colour Developer reacts with the captured antibodies to generate a distinct blue dot at the
location of the control spot and, in the case that HIV-1/HIV-2 antibodies are present in the specimen, a
blue dot also appears at the location of the test spot on the membrane. In the final step, the Clarifying
Solution is then added to the membrane to decrease background colour in order to make the control
and test dots more distinct.
Antigen Selection: The INSTI HIV-1/HIV-2 assay utilizes a combination of recombinant
transmembrane proteins from HIV-1 (gp41) and HIV-2 (gp36). Use of these proteins overcomes
sensitivity and specificity problems associated with tests based on viral lysates or a combination of core
antigen and other viral proteins. 9-13
Antibody Detection: The INSTI HIV-1/HIV-2 assay uses a unique reagent to detect antibodies to HIV-
1/HIV-2. Although primarily designed to detect the IgG class of specific antibodies, the INSTI HIV-1/HIV-
2 assay has been shown to detect antibodies in samples obtained early in infection, during
seroconversion, and low titre anti-HIV-1 samples obtained later in infection.
Test Complexity: The INSTI HIV-1/HIV-2 assay was designed to reduce protocol complexity. The INSTI
HIV-1/HIV-2 assay does not require sample preparation, accurate timing, or several steps, which include
multiple washes and reagents. These requirements increase the complexity of an assay and lead to
procedural errors which may adversely affect sensitivity and specificity. Total test time may vary slightly
depending on specimen type; but results of valid tests are always clearly readable within one minute.
SPECIMEN COLLECTION AND STORAGE
1. For EDTA-whole blood, EDTA-plasma or serum specimens, follow venipuncture blood collection
procedures using lavender-top EDTA anticoagulant tubes (for whole blood and plasma) or red-top
(no anticoagulant) tubes for serum.
2. If plasma or serum is to be used, separate from the blood cells by centrifugation.
3. Serum or EDTA-plasma may be stored at 2-8°C for up to 5 days, stored frozen at ≤ -20°C for 3
months, or stored frozen at ≤ -70°C for one year.
4. Whole blood specimens collected in EDTA anticoagulant may be stored at 2-8°C and should be
tested within 48 hours. Do not heat or freeze whole blood specimens.
5. Do not dilute prior to testing.
KIT COMPONENTS AND STORAGE
INSTI components should be stored at 2-30°C.
For 90-1015 all kit components are individually packaged for single use only. Each test
requires the following materials:
1. Membrane Unit, individually packaged, prepared with control (IgG capture) and test (gp41 and gp36
antigen) reaction spots. For single use only in the INSTI procedure.
2. Sample Diluent Y Solution 1 vial, containing 1.5 ml of tris-glycine buffered solution containing cell
lysis reagents, with adequate space for addition of blood, serum or plasma samples being tested
with INSTI. Ready to use, no mixing or preparation required. Contains 0.1% sodium azide as a
preservative, for single use only in the INSTI procedure. Stable to date and under storage conditions
indicated on label.
3. Colour Developer YSolution 2 vial, containing 1.5 ml of a blue-coloured Borate buffered proprietary
indicator solution designed to detect IgG in the control spot and specific HIV antibodies in the test
spot. For single use only in the INSTI procedure. Ready to use, invert 2-3X immediately before use.
Contains 0.1% sodium azide as a preservative. Stable to date and under storage conditions indicated
on label.
4. Clarifying Solution Y Solution 3 vial, containing 1.5 ml of a proprietary tris-glycine buffered
clarifying solution designed to remove background staining from the membrane unit prior to reading
the INSTI test results. Ready to use, no mixing or preparation required. For single use only in the
INSTI procedure. Contains 0.1% sodium azide as a preservative. Stable to date and under storage
conditions indicated on label.
SUPPORT MATERIALS D
The following materials are required when testing fingerstick whole blood and included with each kit:
1. Single-use Alcohol Swab
2. Single-use Lancet K
3. Single-use Pipette, 50μl
MATERIALS REQUIRED BUT NOT PROVIDED
• Personal protective equipment.
• Appropriate biohazard waste containers and disinfectants.
• Absorbent cotton balls for fingerstick or venipuncture wound closure.
For venipuncture blood collection and testing:
• Venipuncture apparatus if collecting blood samples.
• Appropriate blood collection tubes.
• Appropriate shipping containers.
• Precision pipette capable of delivering 50μl of sample.
MATERIALS AVAILABLE AS AN ACCESSORY TO THE KIT
INSTI HIV-1/HIV-2 Test Controls: Separate HIV-negative human serum substitute and HIV-1/HIV-2
positive de-fibrinated human plasma control samples product no. 90-1036 are available from bioLytical
Laboratories, for use in quality control procedures. Please refer to the section on Quality Control,
following the Assay Procedure, and the INSTI HIV-1/HIV-2 Test Controls Instructions for Use.
WARNINGS
For in vitro diagnostic use only V
It is recommended that the entire Package Insert be read prior to beginning the test procedure. Although
the assay is designed to be simple to use, conformance with the test procedure is necessary to ensure
accurate results.
1. Do not mix reagents from different lots.
2. Do not use reagents or kits beyond the stated expiration date.
3. Do not use the Membrane Unit if the foil pouch has been previously opened or if the packaging
integrity has been compromised. Once the Membrane Unit has been opened, it must be used
immediately.
4. Avoid microbial contamination of reagents.
5. Y Sodium azide is present at 0.1% in all assay reagents. Sodium azide may react with lead or
copper plumbing to form highly explosive metal azides. If products containing sodium azide are
discarded into a drain, flush with large amounts of water to prevent azide build-up. Check with local
regulatory agencies to determine at what concentration sodium azide may cause a product to be
regulated as hazardous waste.
6. The performance characteristics of the INSTI HIV-1/HIV-2 assay have not been established for body
fluids other than EDTA whole blood, fingerstick blood, serum, and EDTA-plasma. The use of blood
collected in anticoagulants other than EDTA has not been validated. Insufficient data are available
7.
8.
9.
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to interpret tests performed on other body fluids, pooled blood or pooled serum and EDTA-plasma,
or products made from such pools.
7. Failure to use the recommended reagent and specimen volumes may result in leakage and/or
overflow of liquids from the membrane unit.
8. If the kit is refrigerated, ensure it is brought to room temperature before performing the test. Use the
INSTI HIV-1/HIV-2 Test Controls to ensure proper kit performance.
9. Y Patients that are on long term antiretroviral drug therapy may give a false negative INSTI HIV-
1/HIV-2 test result.
10. Samples from patients with severe hypogammaglobulinemia conditions such as multiple myeloma
may result in false negative or invalid results with INSTI.
11. Patients with elevated haemoglobin levels may test false negative with INSTI. 15
PRECAUTIONS
1. Y All specimens should be handled as if capable of transmitting infectious diseases. It is
recommended that BioSafety Level 2 practices, or equivalent regulations, be observed. 14
2. Thoroughly wash hands after handling or performing this test.
3. Do not smoke, eat, or drink in areas where specimens or kit reagents are being handled.
4. Wear a lab coat and disposable gloves while handling kit reagents or specimens. Do not pipette by
mouth.
5. Avoid contact with skin and eyes. If contact occurs, wash affected areas with water.
6. Avoid forming aerosols.
7. Y Dispose of all specimens and materials used to perform the test as if they contained infectious
agents. The preferred method of disposal is sterilization by autoclaving for a minimum of one hour
at 121°C followed by incineration. Liquid waste not containing acid and neutralized waste may be
mixed with sodium hypochlorite in volumes such that the final mixture contains 0.5% sodium
hypochlorite (a solution containing 10% household bleach). Allow at least 30 minutes for
decontamination to be completed. Do not autoclave solutions that contain bleach.
8. Spills should be cleaned up and decontaminated in accordance with the user facility’s established
procedures for handling biohazardous spills.
ASSAY PROCEDURE
NOTE: All Membrane Units must be used immediately once opened. All reagents should be
dispensed evenly in the center of the well.
Sampling Fingerstick Blood:
1. Gather support materials (swab, lancet, pipette), one sealed test pouch containing INSTI Membrane
Unit, and one vial each of the Sample Diluent, Colour Developer, and Clarifying Solution for each
test to be performed.
Y CAUTION! The amount of sample (fingerstick blood) is critical. To ensure that the proper
amount of blood is achieved, follow these instructions carefully:
2. Massage the finger to allow the blood to move to the surface (fingertip will become pink). Use heating
pad if available to warm the hand. Hand must be positioned at waist level or lower.
3. Wipe the fingertip with the alcohol swab.
4. As soon as the finger is dry, twist and remove the protective insert from the lancet. Press the finger
firmly at the point just below where the lancet will be applied. With the other hand, place the lancet
on the side of the fingertip and press hard until it clicks. Immediately dispose the used lancet into a
proper sharps container.
5. As the blood droplet forms, hold the pipette horizontally and touch the tip of the pipette to the blood
sample. Capillary action automatically draws the sample to the fill line and stops. If very little blood
trickles out of the puncture, gently apply intermittent pressure below the puncture site to obtain the
required blood volume. If blood is inadequate, perform a second skin puncture using a new lancet.
Sampling EDTA Whole Blood, serum, EDTA-plasma and Test Controls:
1. Bring specimens to room temperature and mix each specimen thoroughly prior to use. Do not heat
or repeatedly freeze/thaw specimens.
2. Gather one sealed test pouch containing INSTI Membrane Unit, and one vial each of the Sample
Diluent, Colour Developer, and Clarifying Solution for each test to be performed.
3. Using a pipette, add 50μl of whole blood, serum, plasma, or kit controls (see Note) to the Sample
Diluent vial. Recap the vial and mix by inversion. Y Adding an excessive amount of specimen may
cause the device to overflow or leak.
NOTE: In POC settings, for INSTI kit controls, it is important to use a 50μl pipette device to add the
control material to the Sample Diluent vial. Do not use the disposable single-use pipette provided for
finger stick blood collection.
General Procedure after Sampling:
1. Tear open the pouch and remove the Membrane Unit without touching the
center well. Place the unit on a level surface. For sample identification
purposes the tab of the Membrane Unit may be labeled with the patient’s
name or number.
NOTE: At this point, it is important that the following steps be
performed immediately and in sequence.
2. Mix the Sample Diluent-specimen mixture by inverting several times
and pour the entire contents to the center of the Membrane Unit well.
(NOTE: Do this within 5 minutes after the specimen has been added to
the Sample Diluent vial). The sample should be absorbed through the
membrane in less than 30 seconds; however, absorption times will vary
slightly depending upon sample type.
3. Resuspend the Colour Developer by slowly inverting to mix the solution
thoroughly until the reagent is evenly suspended and add the entire
contents to the center of the Membrane Unit well. The coloured solution
should flow through completely in about 20 seconds.
4. Open the Clarifying Solution and add the entire contents to the center of
the Membrane Unit well. This will lighten the background colour and facilitate reading. Immediately
read the result while the membrane is still wet. Do not read the results if more than 5 minutes has
elapsed following the addition of Clarifying Solution.
QUALITY CONTROL
Kit Controls:
The INSTI HIV-1/HIV-2 Antibody Test has a built-in IgG capture procedural control that demonstrates
assay validity and adequate sample addition. A blue colour on the control dot indicates that the proper
specimen was added and that the assay procedure was performed correctly. The control dot will appear
on all valid INSTI tests. (Refer to Interpretation of Results, below.)
INSTI HIV-1/HIV-2 Test Controls are available separately for use only with the INSTI HIV-1/HIV-2
Antibody Test. The controls are used to verify test performance and interpretation of results. Kit controls
should be run under the following circumstances:
• for new INSTI operator verification prior to performing testing on patient specimens
• when switching to a new lot number of INSTI test kits
• whenever a new shipment of kits is received
• when temperature during storage of the kit falls outside of 2°-30°C
• when the temperature of the test area falls outside of 15°-30°C
• at regular intervals as determined by the user facility.
Refer to the INSTI HIV-1/HIV-2 Test Controls instructions for use for additional information on the use
of these reagents. It is the responsibility of each user of the INSTI HIV-1/HIV-2 Antibody Test to establish
an adequate quality assurance program to ensure proper performance under their specific locations and
conditions of use.
INTERPRETATION OF RESULTS
• Do not read the results if more than 5 minutes have elapsed following the addition of
Clarifying Solution.
• If using the control samples provided by bioLytical, all Positive Controls must be reactive
with INSTI and all Negative Controls must be non-reactive with INSTI. Controls that produce
incorrect or invalid results must be re-tested with INSTI. If results are still incorrect or invalid,
inform bioLytical Laboratories immediately.
-2
al
ol,
gh
re
Y CAUTION!
Filling is automatic: Never squeeze the tube while sampling.
6. Transfer the blood held in the pipette to the Sample Diluent vial (Solution 1). Align the tip of the
pipette with the Sample Diluent vial and squeeze the bulb to dispense the sample. NOTE: If the
sample will not expel, hold the pipette vertically and slide a finger over (without pressing) the vent
hole, then squeeze the bulb. Recap the vial and mix by inversion. Follow General Procedure after
Sampling, below.
NON-REACTIVE ► One blue dot that is clearly discernable above any
background tint should appear on the membrane. This is the procedural
Control Dot and shows that the test has been performed correctly. The control
dot location is indicated by the letter C printed on the Membrane Unit. No
reaction should be visible at the test spot, located below the control. A nonreactive
result indicates that antibodies to HIV-1/HIV-2 were not detected in
the specimen.
REACTIVE ► Two blue dots that are discernable above any background tint indicate that the specimen
contains HIV-1/HIV-2 antibodies. One dot may be darker than the other. A sample giving this pattern is
considered a preliminary reactive. Following a reactive rapid test result, a venous blood sample must
be drawn in a lavender-top EDTA collection tube (for whole blood or plasma) or red-top tube (for serum),
and forwarded to a laboratory for HIV confirmatory testing.
13

HIV
Instructions for Use
INVALID ► The test is invalid if any of the following occurs:
A. There is no dot on the membrane
B. The test dot appeared without the control dot
C. Uniform tint across the membrane
D. Only blue specks appear on the membrane
A B C D
NOTE: Invalid tests with fingerstick blood samples in POC settings should be repeated with a fresh
sample using a new membrane unit, kit components and support materials. Invalid tests with EDTA
whole blood, EDTA plasma or serum samples in laboratory settings should be repeated using a new
membrane unit and kit components.
INDETERMINATE ► The test is indeterminate if a faint background ring
appeared on the test area. Following an indeterminate INSTI test result, a
venous blood sample must be drawn in a lavender-top EDTA collection tube
(for whole blood or plasma) or red-top tube (for serum), and forwarded to a
laboratory for HIV confirmatory testing.
Please note the following:
1. Following a reactive or indeterminate INSTI test result, a venous blood sample must be drawn in a
lavender-top EDTA collection tube (for whole blood or plasma) or red-top tube (for serum), and
forwarded to a laboratory for HIV confirmatory testing.
2. Depending on the antibody titer, a reactive specimen may be less intense in colour than the
procedural control, or vice versa.
3. Only a blue spot of colour discernibly darker than the background colour should be interpreted as
reactive or positive. In rare instances, a faint background ring may appear around the test spot; this
should not be interpreted as a reactive result. Only tests exhibiting distinct fully formed blue test dot
combined with a distinct fully formed blue control dot should be interpreted as reactive. Colour
intensity may be variable within or between the dots.
4. An invalid result indicates that the test was performed incorrectly or there is a problem with the
sample or device. The absence of a distinct control dot usually indicates that the sample volume was
insufficient. An invalid test must be repeated.
5. A test resulting in a uniform blue tint across the entire membrane, thus obscuring the control and
test spots, can occur when more than 60μL of whole blood is used and the flow through the assay
membrane is obstructed.
6. An individual who has a non-reactive result but was involved in HIV-risk activity is likewise
recommended to obtain additional testing over the next months.
7. To significantly reduce the risk of HIV transmission, it is advisable to refrain from high risk activities
such as unprotected sex and needle sharing at all times.
LIMITATIONS OF THE TEST
• Flow Times
In some instances, samples may exhibit longer than normal flow times (from the time the Sample
Diluent specimen mixture is poured in the membrane well to the time the Clarifying Solution has fully
flowed through the membrane). This is due to variable factors such as cellular components,
especially with whole blood. In instances of long flow times, a faint shadow in the form of a ring
may appear at the test spot location, but this should not be interpreted as a reactive result.
This should be considered as an indeterminate result. In these instances, a venous blood
sample should be drawn in a lavender-top EDTA collection tube, and forwarded to a laboratory for
HIV confirmatory testing.
• The INSTI HIV-1/HIV-2 Antibody Test procedure and the interpretation of result must be followed
closely when testing for the presence of antibodies to HIV in serum, plasma or whole blood.
• Insufficient data are available to interpret tests performed on other body fluids, pooled blood or
pooled serum and plasma, or products made from such pools; therefore, testing of these specimens
is not recommended.
• The INSTI HIV-1/HIV-2 Antibody Test has not been validated for detection of antibodies to HIV-1
Group N subtypes.
• The INSTI HIV-1/HIV-2 Antibody Test detects antibodies to HIV-1/HIV-2 and is useful in establishing
infection with HIV. Because a variety of factors may cause non-specific reactions, a patient found to
be positive using the INSTI HIV-1/HIV-2 assay should have an EDTA blood sample drawn for
laboratory-based confirmatory testing. A person who has antibodies to HIV is presumed to be
infected with the virus and appropriate counseling and medical evaluation should be offered. The
presence of HIV antibodies indicates past exposure to HIV but is not a diagnosis of AIDS, which can
only be made by a physician. However, a non-reactive test does not rule out past exposure to HIV.
The risk of an asymptomatic person with repeated reactive serum developing AIDS is not known.
The prevalence of HIV infection in various groups, as well as clinical and public health guidelines,
are available in the CDC Morbidity and Mortality Report. 8
PERFORMANCE CHARACTERISTICS
SENSITIVITY
DETECTION OF ANTIBODIES TO HIV-1 IN SPECIMENS FROM INDIVIDUALS INFECTED WITH HIV-1
A multi-center prospective study was conducted to evaluate the clinical performance of the INSTI HIV
Antibody Test. There were 483 subjects known to be HIV-1 positive, and 905 subjects with unknown
HIV status. The subjects with unknown HIV status were tested with INSTI and by a composite reference
method (comparator method) which consisted of an licensed/approved EIA with supplemental Western
blot and PCR assays as required. The result of INSTI was compared to the known or determined HIV
status of the subject.
In this study, all 517/517 true HIV antibody positive subjects were identified as reactive by the INSTI
HIV-1/HIV-2 Antibody Test, resulting in a relative sensitivity of 100.0% (95 % CI = 99.3% - 100.0%).
There were no invalid results (0/1388) observed in this study.
Detection of HIV-1 Antibody in Fingerstick Whole Blood Specimens from HIV-1 Seropositive
Individuals
Study Population
Number of
Subjects
INSTI
Reactive
Approved
Test
Reactive
True Positive
HIV status unknown 905 34 34 34
Known HIV-1 Positive 483 483 483 483
TOTAL 1,388 517 517 517
Reactivity with HIV-1: Seroconversion Panels
Thirty (30) HIV-1 seroconversion panels (Boston Biomedica Inc.) were tested with INSTI. Each panel
consisted of sequential serum/plasma specimens obtained from a single individual during
seroconversion. The results of this study are presented in the table below and summarizes the INSTI
HIV-1/HIV-2 assay data compared to US licensed and European approved HIV antibody enzyme
immunoassays (EIA). Overall the INSTI HIV-1/HIV-2 Antibody Test has similar performance to
commercially available anti-HIV EIA in the detection of HIV antibodies in seroconversion samples.
INSTI HIV-1/HIV-2 TEST:
Number of Panels
Detected the earliest bleed that was detected by an EIA 15
Detected within 1 bleed of earliest EIA positive 10
Detected within 2 bleeds of earliest EIA positive 3
Unknown** 2
**The last bleed in the panel was reactive by at least 1 EIA, non-reactive by INSTI
Reactivity with HIV-1: Low Titer Panel
A single low titer HIV-1 antibody panel (#PRB-108; Boston Biomedica) was tested with the INSTI HIV-
1/HIV-2 Antibody Test. This low titer panel consisted of 15 serum/plasma specimens. Results of this
study are summarized in the table below. This study demonstrated that the INSTI HIV-1/HIV-2
Antibody Test has the capability of detecting antibodies to HIV-1 similar to currently available FDA
licensed EIAs.
Specimen Number
Test 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
INSTI HIV-1/HIV-2 P P P P P P P P P P P N P P P
Abbott EIA P P P P P P P P P P P N P P P
Abbott HIVAB HIV-1/HIV2 (rDNA)EIA P P P P P P P P P P P N P P P
Cambridge Biotech Recombigen HIV-1 EIA P P P P P P P P P P P N P P P
Syva EIA P P P P P P P P P P P N P P P
Organon Teknika Vironostika Anti-HIV Uni-
Form II
P P P P P P P P P P P N P P P
Murex HIV 1/2 EIA P P P P P P P P P P P N P P P
Ortho HIV-1/HIV-2 EIA P P P P P P P P P P P N P P P
Sorin ETI-Ab-HIV 1/2K EIA P P P P P P P P P P P N P P P
Syva Microtrak II EIA P P P P P P P P P P P N P P P
Behringwerke ENZ PLUS Anti HIV 1/2 EIA P P P P P P P P P P P N P P P
Biotest Anti-HIV-1/HIV-2 Recombinant EIA P P P P P P P P P P P N P P P
Boehringer Mannheim Anti HIV-1/HIV-2 EIA P P P P P P P P P P P N P P P
IAF Biochem Detect-HIV-EIA P P P P P P P P P P P N P P P
Diagnostic Pasteur Genelavia EIA P P P P P P P P P P P N P P P
bioMerieux VIDAS anti-HIV-1/2 EIA P P P P P P P P P P P N P P P
Murex Wellcozyme HIV-1/HIV-2 EIA P P P P P P P P P P N N P P P
Behringwerke Enzygnost Anti HIV 1+2 EIA N P N P P P P P P P P N P P P
Cellular Products HIV-1 EIA N P P P P P P P N P P N P P P
Genetic Systems LAV EIA N P P P P P P P N P P N P P P
Genetic Systems HIV-1/HIV-2 EIA N P N P P P P P P P P N P P P
*These samples were confirmed positive (P) by EIA and Western Blotting
(Data obtained from Boston Biomedica package insert, May 1995 p.2)
Interfering Substances and Unrelated Medical Conditions
To assess the impact of unrelated medical conditions or interfering substances on the sensitivity of the
INSTI HIV-1/HIV-2 Antibody Test, 195 serum/plasma specimens from a variety of medical conditions
unrelated to HIV-1 infection and 217 specimens with interfering substances were spiked with an HIV-1
positive specimen; see table in the Specificity section for list of medical conditions and substances
tested. All spiked specimens gave reactive results.
DETECTION OF ANTIBODIES TO HIV-2 IN SPECIMENS FROM INDIVIDUALS INFECTED WITH HIV-2
A total of 137 individual HIV-2 positive samples were obtained from European sources. 49 sera from
individuals with chronic HIV-2 infection were reactive on the INSTI HIV-1/HIV-2 Antibody Test. An
additional 88 HIV-2 positive serum and plasma samples were prepared as contrived whole blood; all
88 contrived samples were reactive on the INSTI HIV-1/HIV-2 Antibody test. Combining the results of
the two studies, the relative sensitivity of the INSTI HIV-1/HIV-2 Antibody test for the detection of HIV-
2 antibodies in these studies was calculated to be 100% (137/137).
HIV
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seru
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14

TRAINING PACK
Instructions for Use
e
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-
14 15
P P
P P
P P
P P
P P
P P
P P
P P
P P
P P
P P
P P
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P P
P P
P P
P P
P P
P P
P P
P P
e
1
HIV-1 SUBTYPE TESTING
To assess the sensitivity of the INSTI HIV-1/HIV-2 Antibody test for HIV-1 variants from various
geographic regions, a total of 118 individual confirmed HIV-1 antibody-positive non-B subtype
serum/plasma specimens were tested; of these 118 samples, 109 were non-B subtypes including 23
sub-type O samples. All 118 of these specimens were reactive using INSTI, generating an overall
sensitivity of the INSTI HIV-1/HIV-2 Antibody Test for HIV-1 non-B subtypes of 100%.
SPECIFICITY
A specificity study was performed using 1386 freshly obtained specimens collected from low or unknown
risk and high risk individuals as part of a multicenter prospective clinical trial. Of the 1386 samples,
1376 gave a Non-Reactive result with INSTI and 4 were invalid. INSTI HIV-1/HIV-2 Antibody Test results
were compared to results from a composite reference method (comparator method) which consisted of
an FDA approved EIA with supplemental Western blot and PCR as required. A total of 7 INSTI false
Reactive results (1 from the high risk group, 6 from the low or unknown risk group) were obtained from
the 1382 specimens from HIV-negative individuals that produced valid INSTI results. From this data,
the overall specificity of the INSTI HIV-1/HIV-2 Antibody Test in fingerstick whole blood specimens from
the combined high risk and low or unknown risk populations, minus the invalid results, was calculated
to be 1375/1382 = 99.5% (95% CI = 99.0% - 99.8%).
Performance of the INSTI HIV-1/HIV-2 Antibody Test on Fingerstick Whole Blood Specimens from
Individuals Presumed to be Negative for HIV Infection
Test
Group
Total
Specimens
INSTI
Non-
Reactive 3
Approved
Test Non-
Reactive 2
True
Negative 2
Low Risk 626 620 626 626
High 782 756 1 760 2 760
Risk
TOTAL 1408 1376 1386 1386
1 4 invalid results were not included in the calculation of specificity. The 4 specimens which gave invalid results on INSTI
were Non-Reactive on the approved test.
2 22 Reactives were confirmed by licensed HIV-1 Western Blot and excluded from the calculation of specificity.
3 Of the 22 INSTI Reactive specimens, one was Non-Reactive by the approved test, i.e. INSTI false Reactive.
Interfering Substances and Unrelated Medical Conditions
To asses the impact of unrelated medical conditions or interfering substances on the specificity of the
INSTI HIV-1/HIV-2 Antibody Test, 195 serum/plasma specimens from a variety of medical conditions
unrelated to HIV-1 infection and 217 specimens with interfering substances were analyzed. Five
specimens from individuals with multiple myeloma gave invalid results. No false reactive results were
obtained.
Medical Condition (n=195)
No. of
Specimens
INSTI
Reactive
INSTI
Nonreactive
Toxoplasmosis 20 0 20
Rheumatoid Factor 20 0 20
Multiple Myeloma 10 0 5
Syphilis 30 0 30
SLE 5 0 5
Rubella 20 0 20
Cytomegalovirus 20 0 20
Epstein Barr Virus 20 0 20
HTLV-I/II panel 15 0 15
Hepatitis B Virus 20 0 20
Hepatitis A Virus 15 0 15
Interfering Substances (n=217)
Icteric 20 0 20
Elevated Bilirubin (>8.0mg/dL) 19 0 19
Lipemic 20 0 20
Visual Hemolysis 5 0 5
Elevated Triglyceride (>292mg/dL) 19 0 19
Elevated Hemoglobin (>12g/100mL) 20 0 20
Elevated Albumin (11.5-13.0g/dL) 15 0 15
EDTA 13 0 13
Sodium Heparin 13 0 13
Sodium Citrate 13 0 13
Bacterially Contaminated 60 0 60
In addition, a total of 208 specimens from pregnant women in various trimesters of pregnancy
confirmed to be HIV-1 negative by a 3 rd Generation HIV EIA were tested. One sample (1/208)
produced invalid result, all other INSTI results were non-reactive.
EQUIVALENCE STUDIES
The INSTI HIV-1/HIV-2 Antibody Test was evaluated using matched serum and plasma specimens,
Testing was performed with 50 anti-HIV-1 negative specimens (25 serum and 25 plasma) and 50 anti-
HIV-1-spiked positive specimens. All samples produced acceptable assay performance. These results
indicate 100% relative sensitivity and 100% relative specificity with the matched serum and plasma
panel provided, and that serum and plasma sample types are equivalent.
REPRODUCIBILITY
The reproducibility of the INSTI HIV-1/HIV-2 Antibody Test was tested at 3 laboratory sites using 3 lots
of the INSTI device on 3 separate days. A panel of 9 blind-coded plasma samples, consisting of 4
antibody positive, 1 very low antibody level sample, and 4 antibody negative samples was tested at
each site.
A total of 729 tests were conducted, 243 at each site.
BIBLIOGRAPHY
1. Guyader, M., Emerman, M., Sonigo, P., et al. Genome organization and transactivation of the human
immunodeficiency virus type 2. Nature 326:662-669, 1987.
2. Blattner, W., Gallo, R.C., and Temin, H.M. HIV causes AIDS. Science 241:515, 1988.
3. Curran, J.W., Morgan, W.M., Hardy, A.M., et al. The epidemiology of AIDS; Current status and future
prospects. Science 229:1352-1357, 1985
4. Sarngadharan, M.G., Popovic, M., Bruch, L., Schüpback, J., and Gallo, R.C. Antibodies reactive with
human T-lymphotropic retroviruses (HTLV-III) in the serum of patients with AIDS. Science 224:506-508,
1984
5. Gallo, R.C., Salahuddin, S.Z., Popovic, M., et al. Frequent detection and isolation of cytopathic
retroviruses (HTLV-III) from patients with AIDS and at risk for AIDS. Science 224:500-503, 1984
6. Weber, J.N., Weiss, R.A., Roberts, C., et al. Human immunodeficiency virus in two cohorts of
homosexual men; Neutralising sera and association of anti-gag antibody with prognosis. Lancet 1:119-
124, 1987
7. Clavel, F., Guétard, D., Brun-Vézinet, F., et al. Isolation of a new human retrovirus from West African
patient with AIDS. Science 233:343-346, 1986
8. Centers for Disease Control. Revision of the CDC surveillance case definition for acquired
Immunodeficiency syndrome. MMWR 36 (suppl. no. 1S):1S-15S, 1987
9. World Health Organization/Global Programme on AIDS. Report of a WHO workshop on synthetic
peptides in HIV diagnosis and AIDS-related research, Moscow 24-26 May 1989. WHO Report, AIDS
1991, 5: WHO1-WHO9
10. Los Alamos National Laboratory. Human retroviruses and AIDS Database. A compilation of nucleic
acid and amino acid sequences, 1993.
11. World Health Organization/Global Programme on AIDS. Operational characteristics of commercially
available assays to detect antibodies to HIV-1 and/or HIV-2 in human sera. Geneva, Switzerland: WHO
documents GPA/BMR/89.4; GPA/BMR/90.1; GPA/RES/DIA90.1; GPA/RES/DIA/91.6; GPA/RES/DIA/
92.8 and GPA/RES/DIA/93.4
12. World Health Organization/Global Programme on AIDS. Acquired immunodeficiency syndrome
(AIDS proposed WHO criteria for interpreting results from Western blot assays for HIV-1, HIV-2 and
HTLV-I/HTLVII). WHO Weekly Epidemiological Record 65(37):281-282, 1990
13. Malone, J.D., Smith, E.S., Sheffield, J., et al. Comparative evaluation of six rapid serological tests
for HIV-1 antibody. Journal of Acquired Immune Deficiency Syndrome (JAIDS) 6:115-149, 1993
14. The Laboratory Biosafety Guidelines, 3rd Edition. Office of Laboratory Security, Health Canada.
3.1.2, 2004.
15. “Summary of Safety and Effectiveness Data, 50-1110.” bioLytical Laboratories, Inc. 28 January
2011, Table 3.
TECHNICAL INFORMATION
For further information or assistance, contact the Technical Services at +1 604-644-4677.
Reference herein to any specific third party by name, trade name, trade-mark, manufacturer or otherwise does
not constitute or imply an endorsement or recommendation of this Kit by such third party, or of the products or
services of such third party by bioLytical or that such products or services are necessarily best suited for the
intended purpose.
M Manufactured by:
bioLytical TM Laboratories, Inc.
406- 13251 Delf Place
Richmond, BC
Canada V6V 2A2
Phone: +1 604-204-6784
Fax: +1 604-244-8399
www.biolytical.com
Authorized Representative:
P EMERGO EUROPE
Prinsessegracht 20, 2514 AP, The Hague, The Netherlands
Phone: +31.70.345.8570 Fax: +31.70.346.7299
C
0543
51-1037P 25-Sep-2020
© Copyright 2020, All rights reserved.
-
For the 4 antibody positive and 4 antibody negative samples, the overall reproducibility was 99.7%
(646/648, two antibody negative samples were read as weak positive at 1 site). For the 1 very low level
antibody sample, 59% (48/81) of the results were reactive while 41% (33/81) were non-reactive.
15

HIV
Test Controls IFU
Y
HIV-1/HIV-2 Test Controls
Product Number: 90-1036
Store at -20°C to +8°C H Use by
Caution
Harmful if swallowed M Manufacturer
i Consult Instructions for Use
C CE Mark
C
0543
STORAGE INSTRUCTIONS
1. The INSTI HIV-1/HIV-2 Test Controls are shipped without temperature control.
Upon receipt, store frozen at -20°C (-4°F) for up to one year or refrigerate at 2°C
(36°F) to 8°C (46°F) for up to one year.
2. The shelf life is dictated by the printed expiry date on the INSTI Test Controls.
3. It is recommended to store the vials in an upright position.
4. Once the controls are thawed continue storage at refrigeration temperatures (2-
8°C). They remain stable until expiry (up to 1 year). Do not re-freeze once the
vials have been opened.
5. Fluctuations in temperature causing freeze-thaw cycles may affect the
performance of the INSTI Test Controls.
PROCEDURE
Materials Required but not Provided
• Pipette capable of delivering 50μl of specimen.
• INSTI HIV-1/HIV-2 Antibody Test Instructions for Use.
Read this Instructions for Use and the INSTI ® HIV-1/HIV-2 Antibody Test Instructions for
Use before using this product. Conformance with the test procedure is necessary to
ensure accurate results. Before performing the test, all operators must become familiar
with Universal Precautions for Prevention of Transmission of Human Immunodeficiency
Virus, Hepatitis B Virus, and other Blood-borne Pathogens in Health Care Settings 1
NAME AND INTENDED USE
The INSTI HIV-1/HIV-2 Test Controls are intended to be used only with the INSTI HIV-1/HIV-
2 Antibody Test.
SUMMARY
INSTI HIV-1/HIV-2 Positive and Negative Controls should be used in conjunction with Good
Laboratory Procedures. They should be run under the following circumstances:
• for new INSTI operator verification prior to performing testing on patient specimens
• when switching to a new lot number of INSTI test kits
• whenever a new shipment of kits is received
• when temperature during storage of the kit falls outside of 2°-30°C (35.6°-86°F)
• when the temperature of the test area falls outside of 15°-30°C (59°-86°F)
• at regular intervals as determined by the user facility.
PRINCIPLES OF THE PROCEDURE
The INSTI HIV-1/HIV-2 Test Controls have been designed for use with the INSTI HIV-1/HIV-2
Antibody Test to validate the correct performance of the test procedure in the hands of the
operator.
The INSTI HIV-1/HIV-2 Positive Controls are prepared from inactivated human plasma. It is
negative for HBsAg and anti-HCV by U.S. FDA licensed test procedures.
The Positive Controls have been designed to produce an easily visible but faint blue colour on
the INSTI test spot and a darker blue colour on the control spot.
The INSTI HIV-1/HIV-2 Negative Control is prepared from defibrinated human serum which is
negative for Anti-HIV-1 and Anti-HIV-2, HBsAg, and Anti-HCV. The Negative Control will
produce a blue colour on the procedure control spot, but no colour on the test spot, for a Non-
Reactive INSTI test result.
REAGENTS:
HIV-1 POSITIVE CONTROL
1 vial containing 1.0 ml of inactivated human plasma. Each vial is sufficient for 20 INSTI tests.
The source material has been heat inactivated at 60°C for 60 minutes.
HIV-2 POSITIVE CONTROL
1 vial containing 1.0 ml of inactivated human plasma. Each vial is sufficient for 20 INSTI tests.
The source material has been heat inactivated at 60°C for 60 minutes.
NEGATIVE CONTROL
1 vial containing 1.0 ml of processed human serum substitute, non-reactive for antibodies to
HIV and HCV and non-reactive for HBsAg. Each vial is sufficient for 20 INSTI tests.
WARNINGS & PRECAUTIONS
• For in vitro diagnostic use only.
Safety Precautions:
1. All specimens should be handled as if capable of transmitting infectious agents.
2. Thoroughly wash hands after handling or performing this test.
3. Do not smoke, eat, or drink in areas where specimens or reagents are being
handled.
4. Wear disposable gloves while handling reagents or specimens. Do not pipette by
mouth.
5. Avoid contact with skin and eyes. If contact occurs, wash affected areas with
water.
6. Avoid forming aerosols.
7. Dispose of all specimens and materials used to perform the test in a biohazard
waste container. The preferred method of disposal is sterilization by autoclaving
for a minimum of one hour at 121°C. Disposable materials may be incinerated.
Liquid waste may be mixed with sodium hypochlorite (bleach) in volumes such
that the final mixture contains 1.0% sodium hypochlorite (using a freshly prepared
solution containing 10% household bleach). Allow at least 60 minutes for
decontamination to be completed. Do not autoclave solutions that contain
bleach.
8. Spills should be cleaned up and decontaminated in accordance with the user
facility’s established procedures for handling biohazardous spills.
9. For additional information on bio-safety refer to “Universal Precautions for
Prevention of Transmission of Human Immunodeficiency Virus, Hepatitis B Virus,
and other Blood-borne Pathogens in Health Care Settings” 1 and “Updated U.S.
Public Health Service Guidelines for the Management of Occupational Exposures
to HBV, HCV, and HIV and Recommendations for Postexposure Prophylaxis” 2 .
Instructions for use
1. Read the INSTI HIV-1/HIV-2 Test Controls Instructions for Use prior to using the
INSTI HIV-1/HIV-2 Test Controls.
2. Remove from storage at -20°C (-4°F) to 8°C (46°F) and allow the Controls to reach
room temperature before testing with INSTI. Return Controls to refrigeration
storage at 2-8°C after use.
3. Mix the Controls by swirling before use.
4. Uncap the HIV-1 Positive, HIV-2 Positive or Negative Control vial. Using a 50μl
pipette, collect 50μl of the Control.
5. Transfer the Control sample held in the pipette to the INSTI Sample Diluent vial
(Solution 1). Recap the vial and mix by inversion.
6. Follow the INSTI test procedure as described in the TEST PROCEDURE section of the
INSTI HIV-1/HIV-2 Antibody Test Instructions for Use.
7. All Controls should be tested in the same manner as patient samples.
8. The HIV-1 Positive Control, HIV-2 Positive Control and the Negative Control are
to be run on separate Membrane Units.
INTERPRETATION OF RESULTS
• Follow the interpretation guidelines provided in the INTERPRETATION OF
RESULTS section of the INSTI HIV-1/HIV-2 Antibody Test Instructions for Use.
• Reactive Result: Both the control spot and the test spot show blue colour
development.
• Non-Reactive Result: Only the control spot shows blue colour development.
• Invalid Result: The test is invalid if any of the following occurs:
-There is no blue colour on both the control spot and the test spot
-There is blue colour on the test spot but not on the control spot
-Uniform tint across the membrane
-Only blue specks appear on the membrane
LIMITATIONS OF THE PROCEDURE
The INSTI HIV-1/HIV-2 Test Controls are only validated for use with the INSTI HIV-1/HIV-2
Antibody Test.
1. The TEST PROCEDURE and INTERPRETATION OF RESULTS sections of the
INSTI HIV-1/HIV-2 Antibody Test Instructions for Use must be adhered to when
testing the INSTI HIV-1/HIV-2 Test Controls.
2. Deviations from the procedure outlined in the INSTI HIV-1/HIV-2 Antibody Test
Instructions for Use may produce unreliable results.
3. Do not dilute the INSTI HIV-1/HIV-2 Test Controls. The INSTI HIV-1/HIV-2 Test
Controls are intended for use in undiluted form.
4. Adverse shipping and storage conditions or use of expired reagents may
produce erroneous results.
EXPECTED RESULTS
The HIV-1 Positive Control and HIV-2 Positive Control must be Reactive with INSTI and the
Negative Control must be Non-Reactive with INSTI. Controls that produce incorrect or invalid
results must be re-tested with INSTI.
Contact bioLytical Laboratories’ Technical Support if the INSTI HIV-1/HIV-2 Test Controls do
not produce the expected results.
REFERENCES
1. CDC. Universal precautions for prevention of transmission of human
immunodeficiency virus, hepatitis B virus, and other bloodborne pathogens in
health care settings. MMWR 1988; 37(24):377-388
2. CDC Updated U.S. Public Health Service Guidelines for the Management of
Occupational Exposures to HBV, HCV, and HIV and Recommendations for
Postexposure Prophylaxis. MMWR 2001; 50(RR-11):1-42.
Manufactured by:
bioLytical TM Laboratories, Inc
406- 13251 Delf Place
Richmond, BC
Canada V6V 2A2
Phone: +1 604-204-6784
Fax: +1 604-244-8399
www.biolytical.com
Authorized Representative:
P EMERGO EUROPE
Prinsessegracht 20, 2514 AP
The Hague, The Netherlands
Phone: +31.70.345.8570
Fax: +31.70.346.7299
51-1181E 25-Sep-2020
© Copyright 2020. All rights reserved
Handling Precautions:
1. Do not use INSTI HIV-1/HIV-2 Test Controls beyond the expiration date.
INDICATIONS OF REAGENT INSTABILITY OR DETERIORATION
Positive or Negative Controls that are visibly turbid and/or contain particulate matter should
not be used and should be discarded in accordance with safety precautions.
Please contact the Business Development Manager in your area to request the QA controls.
16

TRAINING PACK
Bibliography
I. Published and Submitted INSTI® Manuscripts, and Reports:
Cook, D., Gilbert, M., DiFrancesco, L., Krajden, M. 2010. “Detection of early sero-conversion HIV infection using
the INSTI ® HIV-1 Antibody Point-Of-Care test.” The Open AIDS Journal 176-179.
This study compared INSTI ® HIV-1 Antibody Point-of-Care (POC) Test to laboratory-based tests for detection
of early sero-conversion (i.e. acute) HIV infections. Of 53 individuals with early HIV infection, the INSTI ® test was
reactive for 34/49 (sensitivity 69.4%; 95%, CI 54.6-81.8%) early-infected individuals whose laboratory-based 3rd
generation HIV EIA test was reactive. In conclusion, the INSTI ® POC test performs well compared with other POC
tests for the detection of early sero-conversion HIV infection, but it may miss 20% to 30% of those detected by
laboratory-based 3rd generation anti-HIV tests. Both POC and laboratory-based anti-HIV tests will fail to detect
a proportion of infected individuals in the first weeks after infection.
Galli, R.A., Green, K.F., La Marca, A., Waldman, L.F. Powers. R.E., Daly, A.C., Shackleton, C.R. 2013. “Evaluation of
the accuracy and ease of use of a rapid HIV-1 Antibody Test performed by untrained operators at the point of
care.” Journal of Clinical Virology 65-69.
This is a prospective study conducted to compare the results of the FDA-cleared, INSTI ® HIV-1 Antibody Test
(INSTI ® ) used by untrained operators on finger-stick whole blood to results obtained by trained laboratory
professionals using FDA-cleared comparator methods (CM) on matching venous blood. Of the 517 HIV positive
subjects (34 new positives and 483 known positives) the concordance between INSTI ® performed by untrained
operators and CM performed by trained laboratory professionals was 100% (95% CI = 99.3–100%). Concordance
for HIV negative results (n = 871) was 99.8% (95%CI = 99.2–99.9%). In conclusion, untrained operators with no
laboratory background were able to perform and interpret the results of INSTI ® on finger-stick blood and limit
of detection specimens with a high degree of accuracy by following only the manufacturer’s written instructions.
Lee B.E., Plitt S., Fenton J., Preiksaitis J.K., Singh A.E. 2011. “Rapid HIV tests in acute care settings in an area of
low HIV prevalence in Canada.” Journal of Virological Methods 66-71.
In this study, rapid HIV testing was performed on serum samples in acute care settings in five hospitals from
urban and rural regions in Alberta, Canada using the INSTI ® HIV-1/HIV-2 Rapid Antibody Test. Parallel standard
HIV antibody tests were performed at the provincial reference laboratory. The study found the sensitivity of the
rapid HIV test compared to standard HIV testing was 100%, specificity was 99.9%, the positive predictive value
was 96% and the negative predictive value was 100%.
Pavie, J., Rachline, A., Loze, B., Niedbalski, L., Delaugerre, C., Laforgerie, E., Plantier, J.C., Rozenbaum, W., Molina,
J.M., Simon, F. 2010. “Sensitivity of five rapid HIV test on oral fluid or finger-stick whole blood: a real-time
comparison in a healthcare setting.” Plos One e11581.
In this study, 200 adults with documented HIV-1 (n = 194) or HIV-2 infection (n = 6) were prospectively screened
with five HIV rapid tests using either oral fluid (OF) or finger-stick whole blood (FSB). 20 randomly selected
HIV-seronegative subjects served as controls, and the results were read blindly. Most patients had HIV-1 subtype
B infection (63.3%) and most were on antiretroviral therapy (68.5%). Sensitivity was 86.5%, 94.5%, 98.5%,
94.9%, 95.8% and 99% respectively, with OraQuick OF, OraQuick FSB, Vikia, Determine, Determine Ag/Ab
Combo and INSTI ® (p<0.0001). In conclusion, rapid HIV tests were less sensitive on oral fluid than on finger-stick
whole blood and less sensitive on finger-stick whole blood than on serum
Singh, A.E., Lee, B., Fenton, J., Preiksaitis, J. 2013. “The INSTI ® HIV-1/HIV-2 antibody test: a review.” Expert Opinion
Medical Diagnostics 299-308.
This article reviews available data on kit characteristics and current performance data on the INSTI ® HIV-1/HIV-2
antibody test are presented together with six other rapid point-of-care tests (RPOCTs) for HIV antibody. The
authors conclude the good performance of INSTI ® HIV-1/HIV-2 antibody test, its ease of use, the rapid availability
of results (< 5 min), and the lack of specialized equipment required to use the kit make this kit a useful addition
to the global market. The unique antigen and flow through technology contained in the kit make it a strong
addition to HIV RPOCTs and to rapid/rapid algorithms used in many resource-limited settings.
17

HIV
Bibliography
WHO Prequalification of Diagnostics Programme. 2013. Product: INSITI HIV-1/HIV-2 Antibody Test.
Number: PQDx 0002-002-00, Geneva: WHO PQDx PR.(Public Report)
This report is a summary of prequalification status for INSTI ® HIV-1/HIV-2 Antibody Test. The test was accepted
for the WHO list of prequalified diagnostics and was listed on 29 August 2013. The WHO outlines the manufacturing,
productions, quality and labeling reviewed and approved for the prequalification. (Add brief statement
on INSTI ® lab evaluation results, which were very good – “In a limited evaluation on a panel of 1079 clinically-derived
specimens, we found an initial sensitivity (95% CI) of 100% (99.1% - 100%) and an initial specificity
(95% CI) of 99.7% (98.9% - 100%) compared to the reference assays. The final sensitivity (95% CI) was 100%
(99.1% - 100%) and the final specificity (95% CI) was 99.7% (98.9% - 100%) compared to the reference assays.
In this study, 0% of the results were recorded as indeterminate. Results were interpreted independently by
three technicians; the inter-reader variability was 0%. The invalid rate was 0%.
18

TRAINING PACK
Clinician’s Briefing Notes
User testimonials
Samedaydoctor is a busy sexual health clinic in London, England. We have been using INSTI ® instant HIV testing
for over six months and in that time have never had a false positive or false negative result. We always confirm
positives by DUO testing and the INSTI ® test is always correct. The test is easy to use, fast and extremely reliable.
Dr Laurence Gerlis, Samedaydoctor
I’m very happy to say that we’ve found the test easy to use; to give reliable results which have on occasion been
verified by independent testing; and to be an effective clear way of eliminating patients HIV concerns.
Dr Séan Cummings MBBS DRCOG DFFP LLM MRCGP, Freedomhealth Ltd.
The INSTI ® HIV rapid POCT kit has revolutionised our private practice. Prior to the test, we were able to offer a
same day HIV testing service for our client group, who are mainly gay men and people who have had some risk
and are now anxious. However, this service was dependent on the client coming into clinic, and during regular
office hours, and involved providing results by telephone, some four hours after the blood was taken, making
managing a reactive or positive result difficult. With the introduction of the INSTI ® test, we have been able to
offer a seven day a week testing service, with appointments into the evening and outside normal office hours.
In addition we are also able to offer a visiting service, due to the portability of the INSTI ® kit, allowing us to
visit patients in their home, place of work or hotel. Having used other rapid HIV POCT tests in my NHS practice,
I have been very impressed with the ease of use and rapid result time offered by the INSTI ® HIV, whilst being
reassured by the true human IgG control, which ensures a gold standard of test validity, which when explained
to patients reassures them of the reliability of the test. I have recommended a number of colleagues to use the
test and recently based a presentation at a national clinical meeting on the practical applications of the test in
expanding innovative testing practice.
Justin Gaffney RN, Consultant Nurse for STI Control/Managing Director Metrosexual Health Limited
At TPA we currently use two kinds of POCT for HIV, the INSTI ® and the Determine 1/2. These tests have enabled
people to come forward for testing and receive a full pre and post- test consultation along with their results all
within the hour. This has encouraged those who would otherwise not have tested due to clinic waiting times
and the length of time it takes to receive a result, to come forward for a check up. The INSTI ® test has a lot
of great benefits as its all packaged together and with using a plastic pipette as opposed to a glass capillary
tube, they have proved to be a safe resource for staff when testing within prison settings and when testing
high risk groups. The reliability of the tests are also a great benefit when reassuring clients of their results. TPA
also took part in a project in Ghana establishing two rapid testing services and when training staff on both
the Determine and the INSTI ® , a high percentage of staff favoured the INSTI ® for its simplicity and speed at
providing a result. I would highly recommend any clinic considering POCT tests to use the test.
Michael Kay, Manager, Teeside Positive Action
19

MULTIPLEX
Four Simple Steps...
The INSTI ® Multiplex HIV-1/2 Syphilis Ab Test is a single-use, rapid, flow-through, in vitro qualitative
immunoassay for the detection of antibodies to Human Immunodeficiency Virus (HIV) Type 1/ Type 2 and
Treponema pallidum in human EDTA-whole blood, fingerstick blood, serum or EDTA-plasma. The test is
intended for use by trained personnel as an in vitro diagnostic device capable of providing results in less than
one minute.
Developed by the same company that brought you the INSTI ® Point of Care HIV test, The INSTI ® Multiplex Test
is an advancement in the testing process that allows the individual to be checked for both HIV and Syphilis.
Just as the original, the test takes as little as 60 seconds for a result. Further advice, help and support can be
given to anyone who tests positive without the need for a lengthy wait.
Add 50μL of ED-
TA-whole blood, fingerstick
blood, serum
or EDTA-plasma with
pipette to the sample
diluent (bottle #1). Close
lid and invert the bottle
(do not shake).
Pour the diluted sample into
the Membrane Unit Well.
HIV-1, HIV-2 and Syphilis
antibodies, if present, are
captured at the test spots.
Add the Colour Developer
(bottle #2) to the center of
the Membrane Unit well to
generate a blue control spot
and up to two additional test
spots if HIV-1 and/or HIV-2
and/or Syphilis antibodies
are present.
Add the Clarifying Solution
(bottle #3) to the Membrane
Unit well to reduce
background colour and
produce a more distinct
test and/or control spot.
• Built-in true human IgG
Control
• Use 50μL of EDTA-whole
blood, fingerstick blood,
serum or plasma
• Rapid results at a glance
(60 seconds)
• Proven early antibody
detection
• Large scale trial data
available
• Easy to understand
instructions
• No timing required
• No refrigeration required
20

TRAINING PACK
Interpretation of Results
HIV SPOT INTERPRETATION
Control
Always read
results with
INSTI ® logo
facing you
HIV
Strong Reactive
Weak Reactive
Indeterminate
Negative
SYPHILIS SPOT INTERPRETATION
Control
Always read
results with
INSTI ® logo
facing you
Syphilis
Strong Reactive
Weak Reactive
Indeterminate
Negative
HIV & SYPHILIS SPOT INTERPRETATION
Control
Always read
results with
INSTI ® logo
facing you
Syphilis
HIV
Strong Reactive
Weak Reactive
21

MULTIPLEX
Instructions for Use
MULTIPLEX
HIV-1/2 Syphilis* Ab Test
*C mark for Syphilis by self-declaration
Single-use rapid assay for the detection of antibodies to
Human Immunodeficiency Virus Type 1 (HIV-1), Type 2 (HIV-2), and T. pallidum
90-1028 - One INSTI ® Multiplex HIV-1/2 Syphilis Ab Test with support materials (for POC use)
Store at 15°C to 30°C
K
Sterilization using irradiation
Caution
Y Harmful if swallowed g
Lot number
V
In Vitro diagnostic medical device
h
Catalogue Number
i Consult instructions for use M Manufacturer
D Do not reuse C CE Mark
H Use by
Store at 15°C – 30°C. For in vitro diagnostic use only.
i It is recommended that the entire Instructions for Use be read prior to beginning the test
procedure. Although the assay is designed to be simple to use, conformance with the test procedure is
necessary to ensure accurate results.
INTENDED USE - Not for donor screening
The INSTI MULTIPLEX HIV-1/2 Syphilis Ab Test is a single use, rapid, flow-through in vitro qualitative
immunoassay for the detection of antibodies to Human Immunodeficiency Virus Type 1/ Type 2 and
Treponema pallidum in human EDTA-whole blood, fingerstick blood, serum or EDTA-plasma. The test is
intended for use by trained personnel in medical facilities, clinical laboratories, emergency care situations,
and physicians’ offices as an in vitro diagnostic device capable of providing results in less than one minute.
It is suitable for near-patient or point-of-care (POC) testing, and is not currently approved for self-testing.
All required pre and post-test counseling guidelines must be followed in each setting in which the INSTI
Multiplex antibody test is used.
The INSTI MULTIPLEX HIV-1/2 Syphilis Ab Test will be referred to as INSTI Multiplex Test in the
remainder of this Instructions for Use.
SUMMARY
Acquired Immunodeficiency Syndrome (AIDS) is caused by at least two retroviruses, HIV-1 and HIV-2.
HIV-1 and HIV-2 are similar in genomic structure, morphology and ability to cause AIDS. 1 HIV is
transmitted mainly by sexual contact, exposure to blood or blood products, or from an infected mother to
her fetus. People with increased risk of HIV infection include haemophiliacs, intravenous drug-users and
men having sex with men (MSM). HIV has been isolated from patients with AIDS, AIDS-related complex
(ARC), and from persons at high risk of contracting AIDS. 2-5 Antibodies specific for HIV envelope proteins
are prevalent in sera from persons at high risk of contracting AIDS as well as in people with AIDS, or
ARC. 5-7 The presence of antibodies to HIV indicated previous exposure to the virus, but does not
necessarily constitute a diagnosis of AIDS. The prevalence of antibodies to HIV in people not known to
be at risk of acquiring HIV infection is unknown, but significantly less. 5 Absence of antibodies to HIV
does not indicate that an individual is free of HIV-1 or HIV-2; HIV has been isolated from
seronegative individuals prior to seroconversion. Test specificity and sensitivity depend, amongst
other factors, on: a) the selection of HIV antigens used for antibody detection, b) the classes of antibodies
recognized by the detection conjugate, and c) complexity of the protocol used to perform the test. 8 Nonspecific
reactions may be observed in some specimens. A reactive INSTI test result should be considered
a preliminary result, with appropriate counseling provided in POC settings. Following a reactive HIV rapid
test result, a venous blood sample must be drawn in an EDTA collection tube (for whole blood or plasma),
and forwarded to a laboratory for HIV confirmatory test.
Treponema pallidum is the causative agent of syphilis. Some of the proteins of this organism are highly
immunoreactive and infected persons develop antibodies soon after infection. These antibodies are
unaffected by treatment and once induced they remain detectable for years. It is possible for a person to
be antibody positive for T. pallidum, but have been cured of the infection. Following a reactive result for
T. pallidum antibodies, a venous blood sample must be drawn in an EDTA collection tube (for whole
blood or plasma) or red-top tube (for serum), and forwarded to a laboratory for syphilis confirmatory
testing. A confirmatory test is required to determine active syphilis or past infection in the patient.
PRINCIPLES OF THE TEST
C0543
for HIV-1/HIV-2 only
The INSTI Multiplex Test is a manual, visually read, flow through immunoassay for the qualitative
detection of HIV-/HIV-2 and syphilis IgG and/or IgM 17 antibodies in human blood, serum or plasma. The
test consists of a synthetic filtration membrane positioned atop an absorbent material within a plastic
cartridge, referred to as the INSTI Membrane Unit. The membrane has been specifically treated with
HIV-1 and HIV-2 recombinant proteins, and syphilis antigens which react with HIV-1/HIV-2 and syphilis
IgG and/or IgM antibodies in the specimen to produce distinct visual signals on the membrane. The
membrane also includes a procedural control. The procedural control consists of a protein-A treated spot
capable of capturing IgG or IgM antibodies normally present in blood and blood components. IgG or IgM
antibodies react with a proprietary chromatic agent to produce a visual signal on the membrane.
Since IgG and/or IgM antibodies can be present in blood from normal or HIV or syphilis positive human
specimens, the control dot provides a visual signal when the test is run, indicating that the test was
performed correctly. If the control dot does not appear, the test is considered invalid. In the case of the
test dots, recombinant HIV-1, HIV-2 and syphilis proteins, embedded in the membrane, capture specific
antibodies, if present in the specimen. Antibodies captured in the test dots react with a proprietary
chromatic agent to produce visible signals on the membrane. The membrane unit is designed to filter,
absorb, and retain the test specimen and all the test reagents in such a manner as to limit leakage and
exposure of personnel to potentially infectious materials.
Reagents required to conduct a test include Sample Diluent, Colour Developer and a Clarifying Solution.
The test is performed by adding the blood, serum or plasma specimen to the vial of Sample Diluent, which
lyses the red blood cells and dilutes the specimens. This specimen/diluent solution is then poured onto
the well of the membrane unit. HIV-1/HIV-2 and syphilis antibodies, if present in the specimen, are
captured by proteins on the filtration membrane. Colour developer is then added to the Membrane Unit.
The Colour Developer reacts with the captured antibodies to generate a distinct blue dot at the location
of the control spot and, in the case that HIV-1/HIV-2 and/or syphilis antibodies are present in the specimen,
a blue dot also appears at the location of one or both of the test spots on the membrane. In the final step,
the Clarifying Solution is then added to the membrane to decrease background colour in order to make
the control and test dots more distinct.
Antigen Selection: The INSTI HIV-1/HIV-2 assay portion utilizes a combination of recombinant
transmembrane proteins from HIV-1 (gp41) and HIV-2 (gp36). Use of these proteins overcomes
sensitivity and specificity problems associated with tests based on viral lysates or a combination of core
antigen and other viral proteins. 9-13 The syphilis antigens bound to the membrane consist of a recombinant
fusion protein derived from p17 and p47 domains of Treponema pallidum.
Antibody Detection: The INSTI Multiplex assay uses a unique reagent to detect antibodies to HIV-1/HIV-
2 and syphilis. Although primarily designed to detect the IgG class of specific antibodies, the INSTI HIV-
1/HIV-2 assay portion has been shown to detect IgM antibodies in samples obtained early in HIV infection
during seroconversion, and low titer anti-HIV-1 samples obtained later in infection 17
Test Complexity: The INSTI Multiplex Test was designed to reduce protocol complexity. The INSTI
Multiplex assay does not require sample preparation, accurate timing, or several steps, which include
multiple washes and reagents. These requirements increase the complexity of an assay and lead to
procedural errors which may adversely affect sensitivity and specificity. Total test time may vary slightly
depending on specimen type but results of valid tests are usually clearly readable within one minute.
SPECIMEN COLLECTION AND STORAGE
1. For EDTA-whole blood, EDTA-plasma or serum specimens, follow venipuncture blood collection
procedures using lavender-top EDTA anticoagulant tubes (for whole blood and plasma) or red-top (no
anticoagulant) tubes for serum.
2. If plasma or serum is to be used, separate from the blood cells by centrifugation.
3. Serum or EDTA-plasma may be stored at 2-8°C for up to 5 days, stored frozen at ≤ -20°C for 3 months,
or stored frozen at ≤ -70°C for one year.
4. Whole blood specimens collected in EDTA anticoagulant may be stored at 2-8°C and should be tested
within 48 hours. Do not heat or freeze whole blood specimens.
5. Do not dilute prior to testing.
KIT COMPONENTS AND STORAGE
INSTI components should be stored at 15-30°C.
All kit components are individually packaged for single use only. Each test requires the
following materials:
1. Membrane Unit, individually packaged, prepared with control (IgG and/or IgM capture), HIV test
(gp41 and gp36 antigen) and T. pallidum (p17-p47 antigen) reaction spots. For single use only in the
INSTI procedure.
2. Sample Diluent, Y Solution 1 vial, containing 1.5 mL of tris-glycine buffered solution containing cell
lysis reagents, with adequate space for addition of blood, serum or plasma samples being tested with
INSTI. Ready to use, invert 2-3X immediately before use.
3. Colour Developer, Y Solution 2 vial, containing 1.5 mL of a blue-coloured borate buffered
proprietary indicator solution designed to detect IgG and IgM in the control spot and specific HIV and
T. pallidum antibodies in the test spots.. Ready to use, invert 2-3X immediately before use.
4. Clarifying Solution, Y Solution 3 vial, containing 1.5 mL of a proprietary tris-glycine buffered
clarifying solution designed to remove background staining from the membrane unit prior to reading
the INSTI test results. Ready to use, no mixing or preparation required
All solutions contain 0.1% Sodium Azide as a preservative and are harmful if swallowed. All solutions are for single
use only and are stable to date and under storage conditions indicated on labels.
SUPPORT MATERIALS
The following materials are required when testing fingerstick whole blood:
1. Single-use Alcohol Swab
2. Single-use Lancet K
3. Single-use Pipette 50μL
MATERIALS REQUIRED BUT NOT PROVIDED
• Personal protective equipment
• Appropriate biohazard waste containers and disinfectants
• Absorbent cotton balls for fingerstick or venipuncture wound closure
For venipuncture blood collection and testing:
• Venipuncture apparatus if collecting blood samples
• Appropriate blood collection tubes
• Appropriate shipping containers
• Precision pipette capable of delivering 50μL of sample
MATERIALS AVAILABLE AS AN ACCESSORY TO THE KIT
INSTI T. pallidum Antibody Positive Control: Separate vials of anti-T. pallidum positive de-fibrinated
human plasma control sample, product no. 90-1032 are available from bioLytical Laboratories.
INSTI HIV-1/HIV-2 Test Controls: Separate HIV-negative human serum substitute and HIV-1/HIV-2
positive de-fibrinated human plasma control samples product no. 90-1036 are available from bioLytical
Laboratories, for use in quality control procedures.
Please refer to the section on Quality Control, following the Assay Procedure, the INSTI Multiplex Test
Controls Instructions for Use and the INSTI HIV-1/HIV-2 Test Controls Instructions for Use.
22

TRAINING PACK
Instructions for Use
WARNINGS
For in vitro diagnostic use only V
It is recommended that the entire Instructions for Use be read prior to beginning the test procedure.
Although the assay is designed to be simple to use, conformance with the test procedure is necessary to
ensure accurate results.
1. Do not mix reagents from different lots.
2. Do not use reagents or kits beyond the stated expiration date.
3. Do not use the Membrane Unit if the foil pouch has been previously opened or if the packaging integrity
has been compromised. Once the Membrane Unit has been opened, it must be used immediately.
4. Avoid microbial contamination of reagents.
5. Y Sodium azide is present at 0.1% in all assay reagents. Sodium azide may react with lead or
copper plumbing to form highly explosive metal azides. If products containing sodium azide are
discarded into a drain, flush with large amounts of water to prevent azide build-up. Check with local
regulatory agencies to determine at what concentration sodium azide may cause a product to be
regulated as hazardous waste.
6. The performance characteristics of the INSTI HIV-1/HIV-2 assay have not been established for body
fluids other than EDTA whole blood, fingerstick blood, serum, and EDTA-plasma. The use of blood
collected in anticoagulants other than EDTA has not been validated. Insufficient data are available to
interpret tests performed on other body fluids, pooled blood or pooled serum and EDTA-plasma, or
products made from such pools.
7. Failure to use the recommended reagent and specimen volumes may result in leakage and/or
overflow of liquids from the membrane unit.
8. If the test kit is exposed to temperatures outside of 15°–30°C, ensure it is brought to this temperature
range before performing testing. Use the Syphilis INSTI Controls and validated HIV Controls to ensure
proper kit performance
9. Y Patients that are on long term antiretroviral drug therapy may give a false negative HIV-1/HIV-2
test result.
10. Samples from patients with severe hypogammaglobulinemia conditions such as multiple myeloma
may result in false negative or invalid results for HIV with INSTI Multiplex.
11. Patients with elevated haemoglobin levels may test false negative for HIV with INSTI Multiplex. 15
12. Because the INSTI Multiplex Test has a lower affinity to IgM antibody class compared to IgG, patients
in the early primary stage of syphilis infection may test negative for T. pallidum antibodies with INSTI
Multiplex.
PRECAUTIONS
1. Y All specimens should be handled as if capable of transmitting infectious diseases. It is
recommended that Directive 2000/54/EC, or equivalent regulations, be observed. 14
2. Thoroughly wash hands after handling or performing this test.
3. Do not smoke, eat, or drink in areas where specimens or kit reagents are being handled.
4. Wear a lab coat and disposable gloves while handling kit reagents or specimens. Do not pipette by
mouth.
5. Avoid contact with skin and eyes. If contact occurs, wash affected areas with water.
6. Avoid forming aerosols.
7. Y Dispose of all specimens and materials used to perform the test as if they contained infectious
agents. The preferred method of disposal is sterilization by autoclaving for a minimum of one hour at
121°C followed by incineration. Liquid waste not containing acid and neutralized waste may be mixed
with sodium hypochlorite in volumes such that the final mixture contains 0.5% sodium hypochlorite (a
solution containing 10% household bleach). Allow at least 30 minutes for decontamination to be
completed. Do not autoclave solutions that contain bleach.
8. Spills should be cleaned up and decontaminated in accordance with the user facility’s established
procedures for handling biohazardous spills.
ASSAY PROCEDURE
NOTE: All INSTI Membrane Units must be used immediately once opened. All reagents should be
dispensed evenly in the center of the well.
Sampling Fingerstick Blood:
1. Gather support materials (swab, lancet, pipette), one sealed test pouch containing INSTI Membrane
Unit, and one vial each of the Sample Diluent, Colour Developer, and Clarifying Solution for each test
to be performed.
Y CAUTION! The amount of sample (fingerstick blood) is critical. To ensure that the proper
amount of blood is achieved, follow these instructions carefully:
2. Massage the finger to allow the blood to move to the surface (fingertip will become pink). Use heating
pad if available to warm the hand. Hand must be positioned at waist level or lower.
3. Wipe the fingertip with the alcohol swab.
4. As soon as the finger is dry, twist and remove the protective insert from the lancet. Press the finger
firmly at the point just below where the lancet will be applied. With the other hand, place the lancet
on the side of the fingertip and press hard until it clicks. Immediately dispose the used lancet into a
proper sharps container.
5. As the blood droplet forms, hold the pipette horizontally and touch the tip of the pipette to the blood
sample. Capillary action automatically draws the sample to the fill line and stops. If very little blood
trickles out of the puncture, gently apply intermittent pressure below the puncture site to obtain the
required blood volume. If blood is inadequate, perform a second skin puncture using a new lancet.
CAUTION!
Filling is automatic: Never squeeze the pipette bulb while sampling.
6. Transfer the blood held in the pipette to the Sample Diluent vial (Solution 1). Align the tip of the
pipette with the Sample Diluent vial and squeeze the bulb to dispense the sample (See Figure A).
NOTE: If the sample will not expel, hold the pipette vertically and slide a finger over (without pressing)
the vent hole, then squeeze the bulb (See Figure B). Recap the vial and mix by inversion. Follow
General Procedure after Sampling, below.
Sampling Whole Blood, serum, plasma and Test Controls:
1. Bring specimens to room temperature and mix each specimen thoroughly prior to use. Do not heat
or repeatedly freeze/thaw specimens.
2. Gather one sealed test pouch containing INSTI Membrane Unit, and one vial each of the Sample
Diluent, Colour Developer, and Clarifying Solution for each test to be performed.
3. Using a pipette, add 50μL of whole blood, serum, plasma, or kit controls (see Note) to the Sample
Diluent vial. Recap the vial and mix by inversion 2-3 times.
Y Adding an excessive amount of specimen may cause the device to overflow or leak.
NOTE: In POC settings, for INSTI kit controls, it is important to use a 50μL pipette device to add the
control material to the Sample Diluent vial. Do not use the disposable single-use pipette provided for
finger stick blood collection.
General Procedure after Sampling:
1. Tear open the pouch and remove the INSTI Membrane Unit without touching the center well. Place
the unit on a level surface. For sample identification purposes the bottom tab of the Membrane Unit
may be labeled with the patient’s name or number.
NOTE: At this point, it is important that the following steps be performed immediately and in
sequence.
2. Mix the Sample Diluent-specimen mixture by inverting several times and pour
the entire contents to the center of the Membrane Unit well. (NOTE: Do this
within 5 minutes after the specimen has been added to the Sample Diluent
vial). The sample should be absorbed through the membrane in less than 30
seconds; however, absorption times will vary slightly depending upon sample
type.
3. Re-suspend the Colour Developer by slowly inverting to mix the solution
thoroughly until the reagent is evenly suspended. Open the Colour Developer
and add the entire contents to the center of the Membrane Unit well. The
coloured solution should flow through completely in about 20 seconds.
4. Open the Clarifying Solution and add the entire contents to the center of the
Membrane Unit well. This will lighten the background colour and facilitate
reading. Immediately read the result while the membrane is still wet. Do not
read the results if more than 5 minutes have elapsed following the
addition of Clarifying Solution.
NOTE: INSTI tests should be read and interpreted under adequate
lighting.
QUALITY CONTROL
A
B
Kit Controls:
The INSTI Multiplex Test has a built-in IgG and IgM capture procedural control that demonstrates assay
validity and adequate sample addition. A blue colour on the control dot indicates that the proper specimen
was added and that the assay procedure was performed correctly. The control dot will appear on all valid
INSTI tests. (Refer to Interpretation of Results, below.)
Separate Syphilis Controls and HIV Controls are available for use with the INSTI Multiplex Test. The
controls are used to verify Syphilis and HIV test performance and interpretation of results. Kit controls
should be run under the following circumstances:
• for new INSTI operator verification prior to performing testing on patient specimens
• when switching to a new lot number of INSTI test kits
• whenever a new shipment of INSTI kits is received
• when temperature during storage of the kit falls outside of 15°-30°C
• when the temperature of the test area falls outside of 15°-30°C
• at regular intervals as determined by the user facility.
Refer to the INSTI T. pallidum Test Controls Instructions for Use and the INSTI HIV-1/HIV-2 Test Controls
Instructions for Use for additional information on the use of these reagents. It is the responsibility of each
user of the INSTI T. pallidum Test Controls to establish an adequate quality assurance program to ensure
proper performance under their specific locations and conditions of use.
CAUTION! It is not recommended to use external controls that have not been validated for the
INSTI Multiplex Test as these may not produce the expected results.
23

MULTIPLEX
Instructions for Use
INTERPRETATION OF RESULTS
• Do not read the results if more than 5 minutes have elapsed following the addition of Clarifying
Solution.
• If using the syphilis control samples provided by bioLytical Laboratories, all syphilis Positive
Controls must be reactive with INSTI and all Negative Controls must be non-reactive with INSTI.
Controls that produce incorrect or invalid results must be re-tested with INSTI. If results are
still incorrect or invalid, inform bioLytical Laboratories immediately.
NON-REACTIVE ► One blue dot that is clearly discernable above any
background tint should appear on the membrane. This is the procedural
Control Dot and shows that the test has been performed correctly. The
control dot is located towards the top of the read frame furthest from the
plastic tab on the Membrane Unit. No reaction should be visible at either of
the two test spots, located below the control. A non-reactive result indicates
that antibodies to HIV-1/HIV-2 and syphilis were not detected in the
specimen.
REACTIVE ► Two or three blue dots that are discernable above any background tint indicate that the
specimen contains HIV-1 and/or HIV-2 and/or syphilis antibodies, depending on the position of the dots.
One dot may be darker than the other. A sample giving these patterns is considered a preliminary reactive.
Following a reactive rapid test result, a venous blood sample must be drawn in an EDTA collection tube
(for whole blood or plasma) or red-top tube (for serum), and forwarded to a laboratory for HIV and/or
syphilis confirmatory testing.
4. An invalid result indicates that the test was performed incorrectly or there is a problem with the sample
or device. The absence of a distinct control dot usually indicates that the sample volume was insufficient.
An invalid test must be repeated.
5. A test resulting in a uniform blue tint across the entire membrane, thus obscuring the control and test
spots, can occur when more than 60μL of whole blood is used and the flow through of the assay
membrane is obstructed.
6. An individual who has a non-reactive result but was involved in HIV-risk activity is recommended to
obtain additional testing over the next months.
7. To significantly reduce the risk of HIV or syphilis transmission, it is advisable to refrain from high risk
activities such as unprotected sex and needle sharing at all times.
LIMITATIONS OF THE TEST
• Flow Times: In some instances, samples may exhibit longer than normal flow times (from the time
the Sample Diluent specimen mixture is poured in the membrane well to the time the Clarifying
Solution has fully flown through the membrane). This is due to variable factors such as cellular
components, especially with whole blood. In instances of long flow times, a faint shadow in the
form of a ring may appear at the test spot location, but this should not be interpreted as a
reactive result. This should be considered as an indeterminate result.
In these instances, a venous blood sample should be drawn in an appropriate collection tube, and
forwarded to a laboratory for HIV and/or syphilis confirmatory testing.
• The INSTI Multiplex Test procedure and the interpretation of result must be followed closely when
testing for the presence of antibodies to HIV and/or syphilis in serum, plasma or whole blood.
• Insufficient data are available to interpret tests performed on other body fluids, pooled blood or pooled
serum and plasma, or products made from such pools; therefore, testing of these specimens is not
recommended.
• The INSTI Multiplex Test has not been validated for detection of antibodies to HIV-1 Group N subtypes.
• The INSTI Multiplex Test detects antibodies to HIV-1/HIV-2 and T. pallidum, and is useful in
establishing infection with HIV and/or syphilis. Because a variety of factors may cause non-specific
reactions, a patient found to be positive for HIV or syphilis using the INSTI Multiplex assay should
have a blood sample drawn for laboratory-based confirmatory testing. A person who has antibodies
to HIV is presumed to be infected with the virus and appropriate counseling and medical evaluation
should be offered. The presence of HIV antibodies indicates past exposure to HIV but is not a
diagnosis of AIDS, which can only be made by a physician. However, a non-reactive test does not
rule out past exposure to HIV. The risk of an asymptomatic person with repeated reactive results
developing AIDS is not known. The prevalence of HIV infection in various groups, as well as clinical
and public health guidelines, are available in the CDC Morbidity and Mortality Report. 8 The presence
of antibodies to T. pallidum may indicate current or past syphilis infection, and a blood sample should
be collected and sent to a laboratory for confirmation of infection status. Antibodies to the syphilis
antigens used in this test may persist for decades, even in spite of successful therapy. A positive
syphilis test may not be an indication of an ongoing infection.
HIV PERFORMANCE CHARACTERISTICS (Note: The HIV-1/HIV-2 portion of the Multiplex assay is
identical to CE Marked INSTI HIV-1/HIV-2 Antibody Test product 90-1015. All data presented in this
section is based on the data presented in the INSTI HIV-1/HIV-2 Antibody Test Instructions for Use,
document 51-1037)
INVALID ► The test is invalid if any of the following occurs:
A. There is no dot on the membrane
B. The test dots appeared without the control dot
C. Uniform tint across the membrane
D. Only scattered blue specks appear on the membrane
SENSITIVITY DETECTION OF ANTIBODIES TO HIV-1 IN SPECIMENS FROM INDIVIDUALS
INFECTED WITH HIV-1
A multi-center prospective study was conducted to evaluate the clinical performance of the INSTI HIV
Antibody Test. There were 483 subjects known to be HIV-1 positive, and 905 subjects with unknown HIV
status. The subjects with unknown HIV status were tested with INSTI and by a composite reference
method (comparator method) which consisted of an licensed/approved EIA with supplemental Western
blot and PCR assays as required. The result of INSTI was compared to the known or determined HIV
status of the subject.
In this study, all 517/517 true HIV antibody positive subjects were identified as reactive by the INSTI HIV-
1/HIV-2 Antibody Test, resulting in a relative sensitivity of 100.0% (95 % CI = 99.3% - 100.0%). There
were no invalid results (0/1388) observed in this study.
Detection of HIV-1 Antibody in Fingerstick Whole Blood Specimens from HIV-1 Seropositive
Individuals
Number
Approved
INSTI
Study Population of
Test True Positive
Reactive
Subjects
Reactive
HIV status unknown 905 34 34 34
Known HIV-1
Positive
483 483 483 483
TOTAL 1,388 517 517 517
NOTE: Invalid tests with fingerstick blood should be repeated with a fresh sample using a new
membrane unit, kit components and support materials. Invalid tests with whole blood, plasma or
serum samples should be repeated using a new membrane unit and kit components.
INDETERMINATE ► The test is indeterminate if a faint background ring appeared on the test areas.
Following an indeterminate INSTI test result, a venous blood sample must be drawn in an EDTA collection
tube (for whole blood or plasma) or red-top tube (for serum), and forwarded to a laboratory for HIV and/or
syphilis confirmatory testing.
Reactivity with HIV-1: Seroconversion Panels
Thirty (30) HIV-1 seroconversion panels (Boston Biomedica Inc.) were tested with INSTI. Each panel
consisted of sequential serum/plasma specimens obtained from a single individual during
seroconversion. The results of this study are presented in the table below and summarizes the INSTI
HIV-1/HIV-2 assay data compared to US licensed and European approved HIV antibody enzyme
immunoassays (EIA). Overall the INSTI HIV-1/HIV-2 Antibody Test has similar performance to
commercially available anti-HIV EIA in the detection of HIV antibodies in seroconversion samples.
INSTI HIV-1/HIV-2 TEST:
Number of Panels
Detected the earliest bleed that was detected by an EIA 15
Detected within 1 bleed of earliest EIA positive 10
Detected within 2 bleeds of earliest EIA positive 3
Unknown** 2
Please note the following:
1. Following a reactive or indeterminate INSTI test result, a venous blood sample must be drawn in an
EDTA collection tube (for whole blood or plasma) or red-top tube (for serum), and forwarded to a
laboratory for HIV and/or syphilis confirmatory testing.
2. Depending on the antibody titer, a reactive specimen may be less intense in colour than the procedural
control, or vice versa.
3. Only a solid blue spot of colour discernibly darker than the background colour should be interpreted as
reactive or positive. In rare instances, a faint background ring may appear around the test spot; this
should not be interpreted as a reactive result. Only tests exhibiting distinct fully formed blue test dot
combined with a distinct fully formed blue control dot should be interpreted as reactive.
**The last bleed in the panel was reactive by at least 1 EIA, non-reactive by INSTI
24

TRAINING PACK
Instructions for Use
Reactivity with HIV-1: Low Titer Panel
A single low titer HIV-1 antibody panel (#PRB-108; Boston Biomedica) was tested with the INSTI HIV-
1/HIV-2 Antibody Test. This low titer panel consisted of 15 serum/plasma specimens. Results of this
study are summarized in the table below. This study demonstrated that the INSTI HIV-1/HIV-2 Antibody
Test has the capability of detecting antibodies to HIV-1 similar to currently available FDA licensed EIAs.
Specimen Number
Test 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
INSTI HIV-1/HIV-2 P P P P P P P P P P P N P P P
Abbott EIA P P P P P P P P P P P N P P P
Abbott HIVAB HIV-1/HIV-2 (rDNA)EIA P P P P P P P P P P P N P P P
Cambridge Biotech Recombigen HIV-1
EIA
P P P P P P P P P P P N P P P
Syva EIA P P P P P P P P P P P N P P P
Organon Teknika Vironostika Anti-HIV
Uni-Form II
P P P P P P P P P P P N P P P
Murex HIV 1/2 EIA P P P P P P P P P P P N P P P
Ortho HIV-1/HIV-2 EIA P P P P P P P P P P P N P P P
Sorin ETI-Ab-HIV 1/2K EIA P P P P P P P P P P P N P P P
Syva Microtrak II EIA P P P P P P P P P P P N P P P
Behringwerke ENZ PLUS Anti HIV 1/2
EIA
Biotest Anti-HIV-1/HIV-2 Recombinant
EIA
Boehringer Mannheim Anti HIV-1/HIV-
2 EIA
P P P P P P P P P P P N P P P
P P P P P P P P P P P N P P P
P P P P P P P P P P P N P P P
IAF Biochem Detect-HIV-EIA P P P P P P P P P P P N P P P
Diagnostic Pasteur Genelavia EIA P P P P P P P P P P P N P P P
bioMerieux VIDAS anti-HIV-1/2 EIA P P P P P P P P P P P N P P P
Murex Wellcozyme HIV-1/HIV-2 EIA P P P P P P P P P P N N P P P
Behringwerke Enzygnost Anti HIV 1+2
EIA
N P N P P P P P P P P N P P P
Cellular Products HIV-1 EIA N P P P P P P P N P P N P P P
Genetic Systems LAV EIA N P P P P P P P N P P N P P P
Genetic Systems HIV-1/HIV-2 EIA N P N P P P P P P P P N P P P
*These samples were confirmed positive (P) by EIA and Western Blotting
(Data obtained from Boston Biomedica package insert, May 1995 p.2)
Interfering Substances and Unrelated Medical Conditions
To assess the impact of unrelated medical conditions or interfering substances on the sensitivity of the
INSTI HIV-1/HIV-2 Antibody Test, 195 serum/plasma specimens from a variety of medical conditions
unrelated to HIV-1 infection and 217 specimens with interfering substances were spiked with an HIV-1
positive specimen; see table in the Specificity section for list of medical conditions and substances
tested. All spiked specimens gave reactive results.
DETECTION OF ANTIBODIES TO HIV-2 IN SPECIMENS FROM INDIVIDUALS INFECTED WITH HIV-
2
A total of 137 individual HIV-2 positive samples were obtained from European sources. 49 sera from
individuals with chronic HIV-2 infection were reactive on the INSTI HIV-1/HIV-2 Antibody Test. An
additional 88 HIV-2 positive serum and plasma samples were prepared as contrived whole blood; all 88
contrived samples were reactive on the INSTI HIV-1/HIV-2 Antibody Test. Combining the results of the
two studies, the relative sensitivity of the INSTI HIV-1/HIV-2 Antibody Test for the detection of HIV-2
antibodies in these studies was calculated to be 100% (137/137).
HIV-1 SUBTYPE TESTING
To assess the sensitivity of the INSTI HIV-1/HIV-2 Antibody Test for HIV-1 variants from various
geographic regions, a total of 118 individual confirmed HIV-1 antibody-positive non-B subtype
serum/plasma specimens were tested; of these 118 samples, 109 were non-B subtypes including 23
sub-type O samples. All 118 of these specimens were reactive using INSTI, generating an overall
sensitivity of the INSTI HIV-1/HIV-2 Antibody Test for HIV-1 non-B subtypes of 100%.
SPECIFICITY
A specificity study was performed using 1386 freshly obtained specimens collected from low or unknown
risk and high risk individuals as part of a multicenter prospective clinical trial. Of the 1386 samples, 1376
gave a Non-Reactive result with INSTI and 4 were invalid. INSTI HIV-1/HIV-2 Antibody Test results were
compared to results from a composite reference method (comparator method) which consisted of an FDA
approved EIA with supplemental Western blot and PCR as required. A total of 7 INSTI false Reactive
results (1 from the high risk group, 6 from the low or unknown risk group) were obtained from the 1382
specimens from HIV-negative individuals that produced valid INSTI results. From this data, the overall
specificity of the INSTI HIV-1/HIV-2 Antibody Test in fingerstick whole blood specimens from the
combined high risk and low or unknown risk populations, minus the invalid results, was calculated to be
1375/1382 = 99.5% (95% CI = 99.0% - 99.8%).
Performance of the INSTI HIV-1/HIV-2 Antibody Test on Fingerstick Whole Blood Specimens from
Individuals Presumed to be Negative for HIV Infection
Test
Group
Total
Specimens
INSTI Non-
Reactive 3
Approved
Test Non-
Reactive 2
True
Negative 2
Low Risk 626 620 626 626
High Risk 782 756 1 760 2 760
TOTAL 1408 1376 1386 1386
1 4 invalid results were not included in the calculation of specificity. The 4 specimens which gave invalid results on INSTI were
Non-Reactive on the approved test.
2 22 Reactives were confirmed by licensed HIV-1 Western Blot and excluded from the calculation of specificity.
3 Of the 22 INSTI Reactive specimens, one was Non-Reactive by the approved test, i.e. INSTI false Reactive.
Interfering Substances and Unrelated Medical Conditions
To assess the impact of unrelated medical conditions or interfering substances on the specificity of the
INSTI HIV-1/HIV-2 Antibody Test, 195 serum/plasma specimens from a variety of medical conditions
unrelated to HIV-1 infection and 217 specimens with interfering substances were analyzed. Five
specimens from individuals with multiple myeloma gave invalid results. No false reactive results were
obtained.
Medical Condition (n=195)
No. of
Specimens
INSTI
Reactive
INSTI
Nonreactive
Toxoplasmosis 20 0 20
Rheumatoid Factor 20 0 20
Multiple Myeloma 10 0 5
Syphilis 30 0 30
SLE 5 0 5
Rubella 20 0 20
Cytomegalovirus 20 0 20
Epstein Barr Virus 20 0 20
HTLV-I/II panel 15 0 15
Hepatitis B Virus 20 0 20
Hepatitis A Virus 15 0 15
Interfering Substances (n=217)
Icteric 20 0 20
Elevated Bilirubin 19 0 19
Lipemic 20 0 20
Visual Hemolysis 5 0 5
Elevated Triglyceride 19 0 19
Elevated Hemoglobin 20 0 20
Elevated Albumin 15 0 15
EDTA 13 0 13
Sodium Heparin 13 0 13
Sodium Citrate 13 0 13
Bacterially Contaminated 60 0 60
In addition, a total of 208 specimens from pregnant women in various trimesters of pregnancy
confirmed to be HIV-1 negative by a 3 rd Generation HIV EIA were tested. One sample (1/208) produced
invalid result, all other INSTI results were non-reactive.
EQUIVALENCE STUDIES
The INSTI HIV-1/HIV-2 Antibody Test was evaluated using matched serum and plasma specimens,
Testing was performed with 50 anti-HIV-1 negative specimens (25 serum and 25 plasma) and 50 anti-
HIV-1-spiked positive specimens. All samples produced acceptable assay performance. These results
indicate 100% relative sensitivity and 100% relative specificity with the matched serum and plasma panel
provided, and that serum and plasma sample types are equivalent.
HIV REPRODUCIBILITY
The reproducibility of the INSTI HIV-1/HIV-2 portion of the Multiplex Test was tested at 3 laboratory sites
using 3 lots of the INSTI HIV-1/HIV-2 Antibody Test on 3 separate days. A panel of 9 blind-coded plasma
samples, consisting of 4 antibody positive, 1 very low antibody level sample, and 4 antibody negative
samples was tested at each site. A total of 729 tests were conducted, 243 at each site. For the 4 antibody
positive and 4 antibody negative samples, the overall reproducibility was 99.7% (646/648, two antibody
negative samples were read as weak positive at 1 site). For the 1 very low level antibody sample, 59%
(48/81) of the results were positive while 41% (33/81) were negative.
SYPHILIS (T. pallidum) PERFORMANCE CHARACTERISTICS
In-house Studies
Data from parallel INSTI Multiplex and T. pallidum Particle Agglutination (TP-PA) in-house testing of
frozen, archived serum and plasma samples obtained from commercial sources is provided below.
INSTI Multiplex T. pallidum antibody test results were compared to a CE marked T. pallidum Particle
Agglutination (TP-PA) assay, n=524 serum/plasma specimens known to be TP-PA negative or positive.
INSTI Multiplex
T. pallidum
Antibody Results
TP-PA Final Interpretation
Positive Negative Total
Reactive 138 5 143
Non-Reactive 7 374 381
Total 145 379 524
NOTE: In an independent study conducted at the Centre National de la Reference de la Syphilis,
Paris, France 16 , the sensitivity of INSTI Multiplex on samples from patients with known secondary
syphilis was 100% (41/41).
Field Study
In an independent prospective field study conducted in Bangalore India in 2012-2013 on a population at
risk for HIV and STI (n=1010 plasma), the performance of the INSTI Multiplex Test was assessed against
the syphilis and HIV status for each patient as determined by an algorithm of syphilis serology methods
(RPR, TPHA) and HIV antibody tests (two rapid lateral flow HIV-1/HIV-2 assays plus one rapid HIV-1/HIV-
2 ELISA test. The results are presented below.
25

MULTIPLEX
Instructions for Use
Syphilis Performance of the INSTI Multiplex Test against the Rapid Plasma Reagin (RPR) test for the
prospective field study population, n=1010.
Rapid Plasma Reagin
(RPR)Test
INSTI Multiplex T. pallidum
Antibody Positive
INSTI Multiplex T. pallidum
Antibody Negative
Positive 12 1 5 2
Negative 2 3 991
Total 14 996
1. 11/12 were confirmed positive by T. pallidum Haemagglutination Assay (TPHA) and 1/12 is considered false positive by
INSTI and RPR.
2. 3/5 were confirmed positive by TPHA, ie INSTI false negative and 2/5 were negative by TPHA, ie RPR biological false
positive.
3. 2/2 were confirmed positive by TPHA, ie RPR false negative
The number of true T. pallidum antibody positive patients (n=16) in this prospective study, as determined
by TPHA is too low for calculation of relative sensitivity of the INSTI Multiplex test for detection of T.
pallidum antibodies. Of the total 994 samples that were considered negative for active syphilis (by RPR
and/or TPHA), INSTI Multiplex was negative for 993, for a negative percent agreement of 99.9%. (It is
important to note that a negative RPR result does not rule out past syphilis infection, therefore this is not
a measure of true T. pallidum antibody specificity.)
HIV Performance of the INSTI Multiplex Test against the HIV test algorithm results for the prospective
field study population, n=1010
Panel
INSTI Multiplex HIV-1/HIV-2
Positive
INSTI Multiplex HIV-1/HIV-2
Negative
HIV known positive (n=136) 136 0
HIV negative (n=874) 0 874
Total 136 874
The positive and negative percent agreement for HIV antibody detection between the INSTI Multiplex
Test and the patients’ HIV status determined by the three-test HIV algorithm was 100%.
Mixed Titre Panel
The INSTI Multiplex Test was tested against a commercial syphilis mixed titre performance panel to
compare performance to other T. pallidum antibody tests for detection of high and low levels of T. pallidum
antibodies. The results are presented below.
Syphilis Mixed Titre Performance Panel 1111-272-00123 (ZeptoMetrix Corporation)
Panel Member ID Number
T. pallidum Test Method 1 2 3 4 5 6 7 8 9 10
US FDA Licensed Test
Pheonix Biotech Trep Sure P P P N N N N N P P
Trinity Captia IgG P P P N N N N N P P
Trinity Captia IgM N N N N N N N N P N
Serodia-TP.PA P P P N N N N N P P
INSTI Multiplex T. pallidum
Antibody
N = negative P = positive
P P P N N N N N P P
Whole Blood Testing
Performance of the INSTI Multiplex Test compared to TP-PA for EDTA Whole Blood Specimens, unspiked
(n=105) and spiked with T.pallidum (n=64) .
INSTI Multiplex
T. pallidum
Antibody Results
TP-PA Final Interpretation
Positive Negative Percent Agreement
Reactive 62 0
Non-Reactive 2 105
Positive percent agreement 96.9%
(62/64)
Negative percent agreement 100%
(105/105)
Total 64 105 169
The TECHNICAL positive percent INFORMATION and negative percent agreement was 96.9% and 100% for this subset of
specimens, NOTE: The comparing HIV assay favorably portion of to the the INSTI corresponding Multiplex Test values is CE obtained Marked from through in-house CE0543 testing and (95.2% the syphilis
and assay 98.7%) portion indicating is CE Marked that there through is no bioLytical performance Laboratories difference by in self-declaration.
the detection of T. pallidum antibodies
in whole blood, serum or plasma samples.
For further information, assistance, or problem reporting, contact Customer Service at +1-604-644-4677.
INSTI Multiplex Test Result compared to TP-PA for specimens that tested positive for other
diseases Reference or medical herein to conditions any specific (n third = 380) party by name, trade name, trade-mark, manufacturer or otherwise
does not constitute or imply an endorsement or recommendation of this Kit by such third party, or of the
products or services of such third party by bioLytical or that such products or services are necessarily
best suited for the intended purpose.
M Manufactured by:
C0543
for HIV-1/HIV-2 only
Condition
No. of
Specimens
1 all were TP-PA positive unless otherwise indicated.
2 all were TPPA negative unless otherwise indicated
3 2 specimens tested negative on TP-PA, ie INSTI false positive
4 2 specimens tested positive on TP-PA, ie INSTI false negative
5 1 specimen tested negative on TP-PA test, ie INSTI false positive
6 1 specimen tested positive on TP-PA, ie INSTI false negative
7 2 specimens tested negative on TP-PA, ie INSTI false positive.
8 3 specimens tested positive on TP-PA test, ie INSTI false negative.
INSTI Multiplex
Syphilis
Reactive 1
INSTI Multiplex
Syphilis
Nonreactive 2
Cytomegalovirus (CMV) 10 1 9
Epstein Barr Virus (EBV) 9 0 9
Helicobactor pylori 10 0 10
Hepatitis A Virus (HAV) 40 6 3 34 4
Hepatitis B Virus (HBV) 40 1 5 39 6
Hepatitis C Virus (HCV) 121 6 7 115 8
Human Immunodeficiency
Virus (HIV)
25 1 24
Herpes Simplex Virus (HSV) 10 0 10
Lyme Disease 5 0 5
Myeloma 10 0 10
Pregnancy 50 0 50
Rheumatoid Factor 5 0 5
Rubella 10 0 10
Systemic Lupus
Erythematosus (SLE)
5 1 4
Toxoplasmosis 20 0 20
Varicella Zoster Virus (VZV) 10 0 10
HIV and Syphilis Reproducibility
The reproducibility of the HIV-1/HIV-2 and syphilis portions of the Multiplex Test was tested using 3
distinct lots of the INSTI Multiplex Test components by 3 operators over 3 separate days. A panel of 5
blind-coded plasma samples, designed to produce HIV and syphilis results ranging from strongly reactive
to weakly reactive to negative, was used for the study. Each panel member sample was tested 33 times,
for a total of 165 INSTI Multiplex tests. For HIV. 165/165 results were in agreement with the expected
results across all operators, component lots and days of testing for an overall reproducibility of 100%.
For syphilis, 164/165 results were in agreement with the expected results across all operators, component
lots and days of testing for an overall reproducibility of 99.4%.
BIBLIOGRAPHY
1. Guyader, M., Emerman, M., Sonigo, P., et al. Genome organization and transactivation of the human
immunodeficiency virus type 2. Nature 326:662-669, 1987.
2. Blattner, W., Gallo, R.C., and Temin, H.M. HIV causes AIDS. Science 241:515, 1988.
3. Curran, J.W., Morgan, W.M., Hardy, A.M., et al. The epidemiology of AIDS; Current status and future
prospects. Science 229:1352-1357, 1985
4. Sarngadharan, M.G., Popovic, M., Bruch, L., Schüpback, J., and Gallo, R.C. Antibodies reactive with
human T-lymphotropic retroviruses (HTLV-III) in the serum of patients with AIDS. Science 224:506-
508,1984
5. Gallo, R.C., Salahuddin, S.Z., Popovic, M., et al. Frequent detection and isolation of cytopathic
retroviruses (HTLV-III) from patients with AIDS and at risk for AIDS. Science 224:500-503, 1984
6. Weber, J.N., Weiss, R.A., Roberts, C., et al. Human immunodeficiency virus in two cohorts of
homosexual men; Neutralising sera and association of anti-gag antibody with prognosis. Lancet
1:119-124, 1987
7. Clavel, F., Guétard, D., Brun-Vézinet, F., et al. Isolation of a new human retrovirus from West African
patient with AIDS. Science 233:343-346, 1986
8. Centers for Disease Control. Revision of the CDC surveillance case definition for acquired
Immunodeficiency syndrome. MMWR 36 (suppl. no. 1S):1S-15S, 1987
9. World Health Organization/Global Programme on AIDS. Report of a WHO workshop on synthetic
peptides in HIV diagnosis and AIDS-related research, Moscow 24-26 May 1989. WHO Report, AIDS
1991, 5: WHO1-WHO9
10. Los Alamos National Laboratory. Human retroviruses and AIDS Database. A compilation of nucleic
acid and amino acid sequences, 1993.
11. World Health Organization/Global Programme on AIDS. Operational characteristics of commercially
available assays to detect antibodies to HIV-1 and/or HIV-2 in human sera. Geneva, Switzerland:
WHO documents GPA/BMR/89.4; GPA/BMR/90.1; GPA/RES/DIA90.1; GPA/RES/DIA/91.6;
GPA/RES/DIA/ 92.8 and GPA/RES/DIA/93.4
12. World Health Organization/Global Programme on AIDS. Acquired immunodeficiency syndrome (AIDS
proposed WHO criteria for interpreting results from Western blot assays for HIV-1, HIV-2 and HTLV-
I/HTLVII). WHO Weekly Epidemiological Record 65(37):281-282, 1990
13. Malone, J.D., Smith, E.S., Sheffield, J., et al. Comparative evaluation of six rapid serological tests for
HIV-1 antibody. Journal of Acquired Immune Deficiency Syndrome (JAIDS) 6:115-149, 1993
14. Directive 2000/54/EC of the European Parliament and of the Council of 18 September 2000 on the
Protection of Workers from Risks Related to Exposure to Biological Agents at Work.
15. “Summary of Safety and Effectiveness Data, 50-1110.” bioLytical Laboratories, Inc. 28 January 2011,
Table 3.
16. Dr Anne Bianchi, CNR, Paris, France, study completed Nov. 2010 / Oct. 2011, personal
communication.
17. N. Moshgabadi et al. Sensitivity of a rapid point of care assay for early HIV antibody detection is
enhanced by its ability to detect HIV gp41 IgM antibodies. Journal of Clinical Virology 71 (2015) 67–
72
TECHNICAL INFORMATION
NOTE: The HIV assay portion of the INSTI Multiplex Test is CE Marked through CE0543 and the syphilis
assay portion is CE Marked through bioLytical Laboratories by self-declaration.
For further information, assistance, or problem reporting, contact Customer Service at +1-604-644-4677.
26
bioLytical TM Laboratories, Inc.
406- 13251 Delf Place
Richmond, BC
Canada V6V 2A2
Phone: +1-604-204-6784
Fax: +1-604-244-8399
www.biolytical.com
51-1143L 25-Sep-2020
©Copyright 2020, All rights reserved.
Authorized Representative:
P EMERGO EUROPE
Prinsessegracht 20, 2514 AP
The Hague, The Netherlands
Phone: +31.70.345.8570 Fax: +31.70.346.7299
Reference herein to any specific third party by name, trade name, trade-mark, manufacturer or otherwise
does not constitute or imply an endorsement or recommendation of this Kit by such third party, or of the
products or services of such third party by bioLytical or that such products or services are necessarily
best suited for the intended purpose.
M Manufactured by:
C0543
for HIV-1/HIV-2 only
bioLytical TM Laboratories, Inc.
406- 13251 Delf Place
Richmond, BC
Authorized Representative:

TRAINING PACK
Syphilis Test Control IFU
Syphilis (T. pallidum) Positive Control
Syphilis Product (T. Number: pallidum) 90-1032 Positive Control
Product Number: 90-1032
Store at -20°C to +8°C H Use by
Store at -20°C to +8°C H Use by
Caution
Y Harmful Caution if swallowed M Manufacturer
Y Harmful if swallowed M Manufacturer
Consult instructions for use
CE Mark
i Consult instructions for use C CE Mark
Read this Instructions for Use and the INSTI ® Multiplex HIV-1/HIV-2/Syphilis Antibody
Test Read Instructions this Instructions for Use for before Use and using the INSTI this product. ® Multiplex Conformance HIV-1/HIV-2/Syphilis with the Antibody test
procedure Test Instructions is necessary for to Use ensure before accurate using this results. product. Before Conformance performing the with test, the all test
operators procedure must is necessary become familiar to ensure with accurate Universal results. Precautions Before performing Prevention the test, of all
Transmission operators must of Human become Immunodeficiency familiar with Virus, Universal Hepatitis Precautions B Virus, and for other Prevention Bloodborne
Transmission Pathogens of in Health Human Care Immunodeficiency Settings 1 . Virus, Hepatitis B Virus, and other Blood-
of
borne Pathogens in Health Care Settings 1 .
NAME AND INTENDED USE
The NAME INSTI AND Syphilis INTENDED (T. pallidum) USE Positive Control is intended to be used only with the INSTI
Multiplex The INSTI HIV-1/HIV-2/Syphilis (T. pallidum) Antibody Positive Test, Control product is number intended 90-1028. to be used only with the INSTI
Multiplex HIV-1/HIV-2/Syphilis Antibody Test, product number 90-1028.
SUMMARY
INSTI SUMMARY Syphilis (T. pallidum) Positive Control should be used in conjunction with Good
Laboratory INSTI Syphilis Procedures. (T. pallidum) It should be Positive run under Control the following should be circumstances:
used in conjunction with Good
• for Laboratory new INSTI Procedures. Multiplex It should operator be run verification under the prior following to performing circumstances: testing on patient
specimens • for new INSTI Multiplex operator verification prior to performing testing on patient
• when specimens switching to a new lot number of INSTI Multiplex test kits
• whenever • switching a new shipment to a new of lot kits number is received of INSTI Multiplex test kits
• when • whenever temperature a new during shipment storage of kits of the is received kit falls outside of 15°- 30°C (59°- 86°F)
• when • when the temperature during of the test storage area of falls the outside kit falls of outside 15°- 30°C of 15°- (59°- 30°C 86°F) (59°- 86°F)
• At • regular when the intervals temperature as determined of the test by area the user falls outside facility of 15°- 30°C (59°- 86°F)
• At regular intervals as determined by the user facility
PRINCIPLES OF THE PROCEDURE
The PRINCIPLES INSTI Syphilis OF (T. THE pallidum) PROCEDURE Positive Control has been designed for use with the INSTI
Multiplex The INSTI HIV-1/HIV-2/Syphilis (T. pallidum) Antibody Positive Test Control to validate has the been correct designed performance use with of the the test INSTI
procedure Multiplex in HIV-1/HIV-2/Syphilis the hands of the operator. Antibody Test to validate the correct performance of the test
procedure in the hands of the operator.
The INSTI Syphilis (T. pallidum) Positive Control is prepared from inactivated human plasma.
It is The negative INSTI for Syphilis HBsAg (T. and pallidum) anti-HCV Positive by U.S. Control FDA licensed is prepared test from procedures. inactivated human plasma.
It is negative for HBsAg and anti-HCV by U.S. FDA licensed test procedures.
The INSTI Syphilis (T. pallidum) Positive Control has been designed to produce an easily
visible The but INSTI faint Syphilis blue colour (T. on pallidum) the INSTI Positive test spot Control and a has darker been blue designed colour on to the produce control an spot. easily
visible but faint blue colour on the INSTI test spot and a darker blue colour on the control spot.
REAGENTS
REAGENTS
SYPHILIS (T. pallidum) POSITIVE CONTROL
1 vial SYPHILIS containing (T. pallidum) 1.0 ml of POSITIVE inactivated CONTROL human plasma. Each vial is sufficient for 20 INSTI
Multiplex 1 vial tests. containing The source 1.0 ml material of inactivated has been human heat inactivated plasma. Each at 60°C vial for is sufficient 60 minutes. for 20 INSTI
Multiplex tests. The source material has been heat inactivated at 60°C for 60 minutes.
WARNINGS & PRECAUTIONS
WARNINGS For & in PRECAUTIONS
vitro diagnostic use only.
For in vitro diagnostic use only.
Safety Precautions:
Safety 1. Precautions:
All specimens should be handled as if capable of transmitting infectious agents.
2. 1. Thoroughly All specimens wash should hands after be handled handling as or if performing capable of transmitting this test. infectious agents.
3. 2. Do Thoroughly not smoke, wash eat, or hands drink after in areas handling where or specimens performing or this kit test. reagents are being
3. handled. Do not smoke, eat, or drink in areas where specimens or kit reagents are being
4. Wear handled. disposable gloves while handling kit reagents or specimens. Do not pipette
4. by mouth. Wear disposable gloves while handling kit reagents or specimens. Do not pipette
5. Avoid by mouth. contact with skin and eyes. If contact occurs, wash affected areas with
5. water. Avoid contact with skin and eyes. If contact occurs, wash affected areas with
6. Avoid water. forming aerosols.
7. 6. Dispose Avoid of forming all specimens aerosols. and materials used to perform the test in a biohazard
7. waste Dispose container. of all The specimens preferred and method materials of disposal used to is perform sterilization the test by autoclaving
a biohazard
for waste a minimum container. of one The hour preferred at 121°C. method Disposable of disposal materials is sterilization may be incinerated. by autoclaving
Liquid for a waste minimum may be of one mixed hour with at sodium 121°C. Disposable hypochlorite materials (bleach) may in volumes be incinerated. such
that Liquid the final waste mixture may contains be mixed 1.0% with sodium sodium hypochlorite hypochlorite (using (bleach) a freshly in volumes prepared such
solution that the containing final mixture 10% contains household 1.0% sodium bleach). hypochlorite Allow at least (using 60 a freshly minutes prepared for
decontamination solution containing to be completed. 10% household Do not bleach). autoclave Allow solutions at least that 60 minutes contain for
bleach. decontamination to be completed. Do not autoclave solutions that contain
8. Spills bleach. should be cleaned up and decontaminated in accordance with the user
8. facility’s Spills established should be procedures cleaned up for and handling decontaminated biohazardous in accordance spills. with the user
9. For facility’s additional established information procedures on bio-safety for handling refer biohazardous to “Universal spills. Precautions for
9. Prevention For additional of Transmission information of Human on bio-safety Immunodeficiency refer to “Universal Virus, Hepatitis Precautions B Virus, for
and Prevention other Blood-borne of Transmission Pathogens of Human in Health Immunodeficiency Care Settings” 1 Virus, and “Updated Hepatitis B U.S. Virus,
Public and Health other Blood-borne Service Guidelines Pathogens for the in Management Health Care of Settings” Occupational 1 and “Updated Exposures U.S.
to HBV, Public HCV, Health and Service HIV and Guidelines Recommendations for the Management for Postexposure of Occupational Prophylaxis” Exposures 2 .
to HBV, HCV, and HIV and Recommendations for Postexposure Prophylaxis” 2 .
Handling Precautions:
Do Handling not use INSTI Precautions: Syphilis (T. pallidum) Positive Control beyond the expiration date.
Do not use INSTI Syphilis (T. pallidum) Positive Control beyond the expiration date.
INDICATIONS OF REAGENT INSTABILITY OR DETERIORATION
INSTI INDICATIONS Syphilis (T. pallidum) OF REAGENT Positive INSTABILITY Control that OR is visibly DETERIORATION
turbid and/or contains particulate
matter INSTI should Syphilis not (T. be used pallidum) and should Positive be Control discarded that in is visibly accordance turbid with and/or safety contains precautions. particulate
matter should not be used and should be discarded in accordance with safety precautions.
STORAGE INSTRUCTIONS
STORAGE 1. The INSTRUCTIONS
INSTI Syphilis (T. pallidum) Positive Control is shipped without temperature
1. control. The INSTI Upon Syphilis receipt, store (T. pallidum) at 2 - 8°C Positive (36 – 46°F) Control for is up shipped to 28 days without or at temperature -20°C (-
4°F) control. for up to Upon 1 year. receipt, store at 2 - 8°C (36 – 46°F) for up to 28 days or at -20°C (-
2. It is 4°F) recommended for up to 1 year. to store the vials in an upright position.
3. 2. Once It is the recommended controls are thawed to store following the vials storage in an upright at -20°C position. (-4°F), they remain stable
3. for Once 28 days the at controls 2 - 8°C are (36 thawed – 46°F). following Do not storage re-freeze at -20°C once (-4°F), the vials they have remain been stable
opened. for 28 days at 2 - 8°C (36 – 46°F). Do not re-freeze once the vials have been
opened.
PROCEDURE
Materials PROCEDURE Required but not Provided
Materials • Precision Required pipette but not capable Provided of delivering 50μl of specimen.
• • INSTI Precision Multiplex pipette HIV-1/HIV-2/Syphilis capable of delivering Antibody 50μl Test of specimen. Instructions for Use.
• INSTI Multiplex HIV-1/HIV-2/Syphilis Antibody Test Instructions for Use.
Instructions for use
Instructions 1. Read for the use INSTI Syphilis (T. pallidum) Positive Control Instructions for Use prior to
1. using Read the the INSTI INSTI Syphilis Syphilis (T. (T. pallidum) pallidum) Positive Positive Control. Control Instructions for Use prior to
2. Remove using the from INSTI storage Syphilis at 2 (T. – 8°C pallidum) (36 – Positive 46°F) or Control. -20 °C (-4°F) and allow the
2. Controls Remove to reach from storage room temperature 2 – 8°C before (36 – testing 46°F) with or -20 INSTI °C (-4°F) Multiplex. and Return allow the
Controls Controls to storage reach at room 2-8°C temperature after use. before testing with INSTI Multiplex. Return
3. Mix Controls the Controls to storage by swirling at 2-8°C before after use. use.
4. 3. Uncap Mix the INSTI Controls Syphilis by swirling (T. pallidum) before use. Positive Control vial. Using a 50μl pipette,
4. collect Uncap 50μl the of INSTI the Control. Syphilis (T. pallidum) Positive Control vial. Using a 50μl pipette,
5. Transfer collect the 50μl Control of the Control. sample held in the pipette to the INSTI Multiplex Sample
5. Diluent Transfer vial (Solution the Control 1). sample Recap the held vial in and the mix pipette by inversion. to the INSTI Multiplex Sample
Diluent vial (Solution 1). Recap the vial and mix by inversion.
6. Follow the INSTI Multiplex test procedure as described in the TEST PROCEDURE
6. section Follow of the INSTI Multiplex test HIV-1/HIV-2/Syphilis procedure as described Antibody in the Test TEST Instructions PROCEDURE for
Use. section of the INSTI Multiplex HIV-1/HIV-2/Syphilis Antibody Test Instructions for
Use.
7. All Controls should be tested in the same manner as patient samples.
8. 7. The All Syphilis Controls Positive should Control, be tested HIV-1 in the Positive same manner Control, as HIV-2 patient Positive samples. Control and
8. the The Negative Syphilis Control Positive are Control, to be run HIV-1 on separate Positive Membrane Control, HIV-2 Units. Positive Control and
the Negative Control are to be run on separate Membrane Units.
INTERPRETATION OF RESULTS
INTERPRETATION • Follow the OF interpretation RESULTS guidelines provided in the INTERPRETATION OF
• RESULTS Follow the section interpretation of the INSTI guidelines Multiplex provided HIV-1/HIV-2/Syphilis in the INTERPRETATION Antibody Test OF
Instructions RESULTS for section Use. of the INSTI Multiplex HIV-1/HIV-2/Syphilis Antibody Test
• Reactive
Instructions
Result:
for
Both
Use.
the control spot and the syphilis test spot show blue colour
• development. Reactive Result: Both the control spot and the syphilis test spot show blue colour
• Non-Reactive development. Result: Only the control spot shows blue colour development.
• • Invalid Non-Reactive Result: The Result: test is Only invalid the if control any of the spot following shows blue occurs: colour development.
• -There Invalid is no Result: blue colour The test on both is invalid the control if any of spot the and following the test occurs: spot
-There -There is blue is no colour blue colour on the on test both spot the but control not on spot the control and the spot test spot
-Uniform -There tint is blue across colour the membrane on the test spot but not on the control spot
-Only -Uniform blue specks tint across appear the on membrane the membrane
-Only blue specks appear on the membrane
LIMITATIONS OF THE PROCEDURE
The
LIMITATIONS
INSTI Syphilis
OF
(T.
THE
pallidum)
PROCEDURE
Positive Control is only validated for use with the INSTI
Multiplex The INSTI HIV-1/HIV-2/Syphilis (T. pallidum) Antibody Positive Test. Control is only validated for use with the INSTI
Multiplex
1. The
HIV-1/HIV-2/Syphilis
TEST PROCEDURE
Antibody
and INTERPRETATION
Test.
OF RESULTS sections of the
1. INSTI The Multiplex TEST PROCEDURE HIV-1/HIV-2/Syphilis and INTERPRETATION Antibody Test instructions OF RESULTS for use sections must be of the
adhered INSTI to Multiplex when testing HIV-1/HIV-2/Syphilis the INSTI Syphilis Antibody (T. pallidum) Test instructions Positive Control. for use must be
2. Deviations
adhered
from
to when
the procedure
testing the
outlined
INSTI Syphilis
in the INSTI
(T. pallidum) ® Multiplex
Positive
HIV-1/HIV-
Control.
2. 2/Syphilis Deviations Antibody from the Test procedure Instructions outlined for Use in may the INSTI produce ® Multiplex unreliable HIV-1/HIV- results.
3. Do
2/Syphilis
not dilute the
Antibody
INSTI
Test
Syphilis
Instructions
(T. pallidum)
for Use
Positive
may produce
Control.
unreliable
The INSTI
results.
3. Syphilis Do not (T. dilute pallidum) the INSTI Positive Syphilis Control (T. is pallidum) intended Positive for use Control. in undiluted The form. INSTI
4. Adverse
Syphilis
shipping
(T. pallidum)
and storage
Positive
conditions
Control is
or
intended
use of expired
for use
reagents
in undiluted
may
form.
4. produce Adverse erroneous shipping results. and storage conditions or use of expired reagents may
produce erroneous results.
EXPECTED RESULTS
The
EXPECTED
INSTI Syphilis
RESULTS
(T. pallidum) Positive Control must be Reactive with INSTI Multiplex.
The INSTI Syphilis (T. pallidum) Positive Control must be Reactive with INSTI Multiplex.
Contact bioLytical Laboratories’ Technical Support if the INSTI Syphilis (T. pallidum) Positive
Control Contact does bioLytical not produce Laboratories’ the expected Technical results. Support if the INSTI Syphilis (T. pallidum) Positive
Control does not produce the expected results.
REFERENCES
REFERENCES
1. CDC. Universal precautions for prevention of transmission of human immunodeficiency
1. virus, CDC. hepatitis Universal B virus, precautions and other for bloodborne prevention pathogens of transmission in health of human care settings. immunodeficiency MMWR
1988; virus, 37(24):377-388 hepatitis B virus, and other bloodborne pathogens in health care settings. MMWR
1988; 37(24):377-388
2. CDC Updated U.S. Public Health Service Guidelines for the Management of Occupational
2. Exposures CDC Updated to HBV, U.S. HCV, Public and Health HIV and Service Recommendations Guidelines for for the Postexposure Management Prophylaxis. of Occupational
MMWR Exposures 2001; to 50(RR-11):1-42.
HBV, HCV, and HIV and Recommendations for Postexposure Prophylaxis.
MMWR 2001; 50(RR-11):1-42.
Manufactured by:
Authorized Representative:
Manufactured by:
Authorized Representative:
EMERGO EUROPE
EMERGO EUROPE
Prinsessegracht 20, 2514 AP
The Prinsessegracht Hague, The Netherlands 20, 2514 AP
bioLytical TM Laboratories, Inc
Phone: The +31.70.345.8570
Hague, The Netherlands
406-13251 bioLytical Delf TM Laboratories, Place Inc
Fax: Phone: +31.70.346.7299 +31.70.345.8570
Richmond, 406-13251 BC, Delf V6V Place
Fax: +31.70.346.7299
2A2 Canada
Phone: Richmond, +1 604-204-6784 BC, V6V 2A2 Canada
Fax: Phone: +1 604-244-8399 +1 604-204-6784
51-1169G 25-Sep-2020
www.biolytical.com
Fax: +1 604-244-8399
© Copyright 2020. 51-1169G All rights 25-Sep-2020
reserved
www.biolytical.com
© Copyright 2020. All rights reserved
27

SELF TEST
Introduction
What is an INSTI ® HIV Self Test
INSTI ® HIV Self Test is designed as a single-use HIV test kit that allows you to get results in the comfort of your
home and at a time that works best for you. Sample, pour and read your results immediately. It’s that easy.
INSTI is designed to be a simple and straightforward home HIV test.
Read the step-by-step instructions inside the box to learn how to use your test and get the results you need to
take control of your health. You can visit www.hivhometest.com to watch a training video.
In-Store Stockist Support:
New Stockist Pack including Posters, Shelf Wobblers & Leaflets to help promote
the product
Online portal of resources
Online Stockist Support:
Online portal of resources to promote the product on your online platform
How to use video which can be hosted on your website or linked to the
product website
Easy to Use
A quick and easy
blood test with three
simple steps
Sensitive
Can detect HIV up to
2 weeks sooner than
other brands
Unique
Novel flowthrough
platform
Accurate
Over 99%
accurate
Clear
Simple dots for
easy interpretation
The World’s Fastest HIV Test
Get highly accurate results
instantly.
Easy to Use
Simple design. No timers,
no fuss.
Early Detection
Detects HIV infection up
to two weeks sooner than
other leading brands.
Convenient and Discreet
Get results from the privacy
of your own home.
28

®
HI V- 1/HI V- 2 Antibody T es t
For in vitro diagnostic use only.
30°C Store at 15-30°C
15°C
51-1061D
Step 2
TRAINING PACK
Instructions For Use
This Test Contains:
SHAKE
4 TIMES
HIV Self Test
Instructions For Use
READ BEFORE USE
Bottle 1, 2, 3
Lancet
STERILE R
Membrane Unit
Test
Device
Bandage
1. S
W
Preparation
Training video available at: www.INSTI.com
1. Open test device pouch.
IMPORTANT: Clean and dry hands.
Step 1: Collect Blood
If you have trouble collecting blood, see Frequently Asked Questions on reverse side.
2. Place test device on a flat surface. 3. Remove cap of Bottle 1. Place on
flat surface.
WARNING: Bottle 1 contains liquid. Handle with care.
Y
PRESS HARD
1. Twist and pull out lancet tip. Discard tip.
2. Rub finger and hand to increase blood flow. 3. Place lancet on the side of finger tip.
As w
th
a
yo
y
4. Rub finger to create a
LARGE drop of blood.
5. Let 1 drop FALL into Bottle 1.
Twist on cap of Bottle 1.
6. Apply adhesive bandage.
Dispo
Dispose
with loc
Put all it
the oute
Throw a
waste bi
29

SELF TEST
Instructions For Use
Step 2: Test
SHAKE
4 TIMES
POUR ALL SHAKE POUR ALL SHAKE POUR ALL
4 TIMES
4 TIMES
1. Shake and pour all liquid.
Wait until liquid disappears.
2. Shake and pour all liquid.
Wait until liquid disappears.
TIP: You may need to gently tap Bottle 2 to get all the liquid out.
3. Shake and pour all liquid.
Wait until liquid disappears.
Step 3: Read Result
Read result right away to within 1 HOUR.
CONTROL
TEST
on
re.
Negative Positive Invalid
Your test result is negative.
Two dots means your test result
is positive. You are probably HIV
positive. Positive results MUST be
confirmed by a doctor.
Your test did not work.
Control dot must appear to
indicate that the test has
been performed correctly.
ARD
TIP: One dot may be lighter than the other. In rare instances, a faint ring may appear at the test dot; this is a positive result.
r tip.
A Negative Result
As with many tests, there is a chance for false results. To reduce
the chance of false results, be sure to follow the instructions
and use the test correctly. If you have a negative result but
you were involved in an HIV-risk activity in the past 3 months,
you could be in what is called the “window period” and it is
recommended to repeat testing at a later date.
A Positive Result
Consult a doctor as soon as possible and inform him/her that
you have performed a self test for HIV. All positive results must
be confirmed by a laboratory test.
What Next After A Positive Result?
Having HIV does not mean you have AIDS. With early diagnosis
and treatment, it is unlikely that you will develop AIDS.
Disposal
Dispose in accordance
with local regulations.
Put all items back into
the outer packaging.
Throw away into
waste bin.
51-1200D 25-Sep-2020
30

TRAINING PACK
Instructions For Use
Store at 2 to 30°C
Caution
Instructions for Use
90-1041 INSTI ® HIV Self Test, Pouch Format
90-1050 INSTI ® HIV Self Test, Box Format
K Sterilization using irradiation
Y
Harmful if swallowed. g Lot number
V In Vitro diagnostic medical device
h
Catalogue Number
i Consult Instructions for Use M Manufacturer
D
H
Do not reuse
Use by
For in vitro diagnostic use only. V
C
P
CE Mark
Authorized Representative in the
European Community
i Read this Instructions for Use prior to beginning the test procedure. Although the INSTI HIV
Self Test is designed to be simple to use, conformance with the test procedure is necessary to
ensure accurate results.
BACKGROUND
HIV stands for Human Immunodeficiency Virus. HIV is the virus that causes AIDS (Acquired
Immunodeficiency Syndrome) if left untreated. It is estimated that there are over 30 million people
living with HIV in the world today, and up to half of those people have not been diagnosed and are
unaware of their infection. This undiagnosed population accounts for most of the HIV transmissions
worldwide. Treatment for HIV is highly effective. It is important to start treatment as early as possible
following infection, to reduce the risk of serious illness or death.
INTENDED USE
The INSTI HIV Self Test is a single use, rapid, flow-through in vitro qualitative immunoassay for the
detection of antibodies to Human Immunodeficiency Virus Type 1 (HIV-1) and Type 2 (HIV-2) in human
fingerstick whole blood. The test is intended for use by untrained lay users as a self test to aid in the
diagnosis of HIV-1 and HIV-2 infection using a small drop (50µL) of blood obtained through fingerstick
collection procedures.
BIOLOGICAL PRINCIPLES OF THE TEST
The INSTI HIV Self Test is an immunoassay for detecting HIV antibodies. The test device consists of
a synthetic membrane positioned atop an absorbent pad within a plastic cartridge. One section of the
membrane has been treated with non-hazardous HIV-1 and HIV-2 recombinant proteins, which capture
HIV antibodies (test dot). The membrane also includes a control dot treated with protein-A that captures
other non-specific antibodies normally present in human blood. The test is performed by adding a
fingerstick blood sample to Bottle 1. The diluted blood in Bottle 1 is poured into the well of the test
device. Any HIV antibodies in the sample are captured by the test dot and non-specific antibodies are
captured by the control dot. Bottle 2 is then added to the test device. Bottle 2 solution reacts with
captured antibodies to produce a blue dot at control dot and, if HIV antibodies are present, a blue dot
also appears at test dot. In the final step, Bottle 3 is added to the membrane to make the control and
test dots more visible.
MATERIALS PROVIDED
1 Instructions for Use
1 Pouch with test device (labelled Membrane Unit)
1 Sample Diluent (Bottle 1, red cap)
1 Colour Developer (Bottle 2, blue cap )
1 Clarifying Solution (Bottle 3, clear cap )
1 Sterile single-use lancet
1 Adhesive bandage
1 Resources Card
C
0543
Test device (packaged inside the pouch labelled Membrane Unit)
The control and/or test dot will appear on the test device once the test is performed. The test device is
prepared with control (IgM/IgG capture) and test (gp41 and gp36 antigen) reaction dots. It is individually
packaged and is to be used only once to complete a single INSTI test.
Sample Diluent (Bottle 1, red cap) Y
A solution designed to dilute the blood sample and break down red blood cells. It contains 1.5 mL of
colorless Tris-Glycine buffered solution containing cell lysis reagents.
Color Developer (Bottle 2, blue cap) Y
A blue solution that detects human antibodies. It contains 1.5 mL of a blue-coloured Borate buffered
proprietary indicator solution designed to detect IgM/IgG in the control dot and HIV-specific antibodies
in the test dot.
Clarifying Solution (Bottle 3, clear cap) Y
A solution to remove background blue color. It contains 1.5 mL of a colorless Tris-Glycine buffered
solution designed to remove background staining from the test device prior to reading the INSTI test
results.
All solutions contain 0.1% Sodium Azide as a preservative and are harmful if swallowed. All solutions
are for single use only and are stable to date and under storage conditions indicated on labels.
LIMITATIONS OF THE TEST
• In some instances, samples may exhibit longer than normal flow times from the time the diluted
blood-Bottle 1 mixture is poured into the test device, to the time the contents of Bottle 3 have fully
flowed through the test device. This is due to variable factors, such as cellular components within
the whole blood sample.
• The INSTI HIV Self Test procedure and the interpretation of results must be followed closely when
testing for the presence of antibodies to HIV.
• This test has not been validated for detection of antibodies to HIV-1 Group N subtypes.
• Because a variety of factors may cause non-specific reactions, a patient found to be positive using
the INSTI HIV Self Test must have the results confirmed by a doctor.
• The presence of HIV antibodies indicates past exposure to HIV but is not a diagnosis of AIDS,
which can only be made by a physician.
• A negative test result does not rule out exposure to HIV.
WARNINGS AND PRECAUTIONS
• Each device is for single use only and is designed for self testing by one person. V
• Keep out of the reach of children.
• The test is for use only with human whole blood.
• Do not use the INSTI HIV Self Test beyond the expiration date stated on the outer packaging.
• Do not use the test device if the foil pouch has been damaged.
• Wash your hands with warm water and ensure they are clean and dry before beginning the test.
• Do not read the result if more than 1 hour has passed after completing the test procedure.
• Failure to follow the instructions may result in leakage and/or overflow of liquids from the test device.
• If you have been on long term antiretroviral drug therapy your test may give a false negative result.
• If you have a severe blood disorder such as multiple myeloma you may obtain a false negative or
invalid result.
• If you have higher than normal haemoglobin, you may test false negative.
• Y All blood samples should be handled as if capable of transmitting infectious diseases.
• Spills should be cleaned up with household bleach or disinfecting wipes.
• Solutions in Bottle 1, 2 and 3 are harmful if swallowed due to the presence of Sodium Azide.
• Test procedure must be completed in the proper sequence without delays between steps.
• Adequate lighting is required to read the test results.
Restrictions on Use
• Not suitable for users who are afraid of needles
• May not be suitable for patients who have been infected within the last 3 months
• Not suitable for users who have a bleeding disorder
• Not suitable for users below the age of 18
• Not suitable for users who are taking anti-retroviral treatment (ART)
• Not suitable for users who have participated in a HIV vaccine study
Storage
• Store in the original packaging in a cool, dry location between 2 to 30°C. DO NOT FREEZE.
• Do not store near a heat source or in direct sunlight.
• The test should be performed at room temperature (15 to 30°C).
• Do not open the test device pouch until you are ready to perform the test. Note that although the
test device pouch states storage at 15-30°C, it can be stored refrigerated, if required.
Disposal
Used safety lancets might be classified as medical waste by health authorities in your area. To reduce
the risk of injury from a used lancet, please follow local requirements for its disposal. Consult your
pharmacist.
Put all other items back into the outer packaging. Throw away into waste bin. Dispose in accordance
with local regulations.
PERFORMANCE CHARACTERISTICS
DIAGNOSTIC SENSITIVITY
Diagnostic sensitivity of a test like the INSTI HIV Self Test is a measure of how well the test detects
the presence of HIV antibodies. Sensitivity is expressed as a percentage and is calculated from data
from a clinical trial or performance evaluation. Sensitivity is calculated by dividing the number of INSTI
positive test results by the number of truly HIV positive persons tested. The higher the sensitivity the
better the test is at correctly identifying truly infected persons. In a study conducted by untrained lay
users (Table 1), 517/517 true HIV antibody positive subjects were identified as positive by the INSTI
test, resulting in a relative sensitivity of 100%.
Table 1- Relative Sensitivity and Specificity of the INSTI Self Test compared to the HIV Status
of Individuals with Known and Unknown HIV Status by Untrained Lay Users
Study
Population
HIV status
unknown
Known HIV-1
Positive
Number of
Subjects
905
483
Total 1,388
Percent of invalid results was 0% (0/1388)
Relative
Sensitivity
100%
(34/34)
100%
(483/483)
100%
(517/517)
95% Confidence
Interval
89.9% - 100%
Relative
Specificity
99.8%
(869/871)
95%
Confidence
Interval
99.2% -
99.9%
99.2% -100% N.A. N.A.
99.3% - 100%
99.8%
(869/871)
99.2% -
99.9%
Studies to calculate the HIV-2 sensitivity of the INSTI HIV Self Test
The sensitivity of INSTI HIV-1/HIV-2 Antibody Test evaluated in an independent European study with
49 sera from Western Blot confirmed HIV-2 infected patients at the chronic stage of the infection was
100%.
Additional studies conducted in-house with 88 different HIV-2 positive serum and plasma samples
obtained from European sources and 24 different plasma samples obtained from Nigeria added into
individual whole blood (to simulate HIV-2 positive blood) also showed 100% sensitivity of INSTI for
HIV-2 antibody detection.
Table 2- INSTI HIV-1/HIV-2 Antibody Test’s Sensitivity in HIV-2 Positive Specimens
Sample Source France 1 France 2 Nigeria 3 Total
Positive Samples 49 88 24 161
INSTI Positives 49 88 24 161
Sensitivity 100% 100% 100% 100%
1. Tests performed in France using serum samples
2. Tests performed in-house using whole blood spiked with plasma (13 samples) and serum (75
samples)
3. Tests performed in-house using plasma samples
In addition, 12 out of 500 prospectively collected plasma specimens from an HIV-2 endemic region
(Ivory Coast) were tested and confirmed as HIV-2 true positive by an FDA-approved differentiation
assay or an HIV-2 RNA quantitative assay.
INSTI was reactive in all 12 of these specimens for a sensitivity of 100%. Results are summarized in
Table 3.
Table 3-
Individua
Spe
End
1 Determin
DIAGNO
Diagnosti
healthy p
data from
INSTI ne
the speci
Self Test
This mea
A specific
individual
and 4 we
specimen
was calcu
1000 test
Table 4-
Specime
Infection
Test Gro
Low or
Unknown
Risk
High Ris
Total
1
Invalid r
invalid re
2
Reactive
specificity
3 Of the 2
Reactive.
Untraine
The perfo
sites. At
experienc
from a tot
tested wi
The sens
invalid re
Unrelate
To asses
sensitivity
condition
tested “un
of reactiv
performa
1. Up to
depen
2. Of the
2 out
Reprodu
The repro
results w
(3 per site
three low
tests were
interprete
of 100% (
FREQUE
What is H
HIV stan
Immunod
begins to
that perso
When his
no cure fo
How doe
HIV sprea
milk of an
to blood t
their babi
HIV throu
HIV cann
infection
a person
from an in
with HIV.
How do I
Relax an
washing
promote b
and not c
What is a
31

SELF TEST
Instructions For Use
.
sing
IDS,
st.
vice.
sult.
e or
the
duce
your
ance
tects
data
STI
y the
d lay
STI
atus
%
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9%
A.
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ation
ed in
Table 3- Detection of Antibody to HIV-2 in Specimens from HIV-2 Seropositive Individuals and
Individuals from an HIV-2 Endemic Region
Specimen Group
Total
Specimens
HIV-2 Positive
INSTI HIV-1/HIV-2
Reactive
Endemic subjects 500 12 1 12
1 Determined by an approved HIV-1/HIV-2 differentiation assay or HIV-2 RNA testing
DIAGNOSTIC SPECIFICITY
Diagnostic specificity of a test like the INSTI HIV Self Test is a measure of how well the test detects
healthy patients who do not have HIV. Specificity is expressed as a percentage and is calculated from
data from a clinical trial or performance evaluation. Specificity is calculated by dividing the number of
INSTI negative test results by the number of truly HIV negative persons that were tested. The higher
the specificity the better the test is at correctly identifying truly non-infected persons. The INSTI HIV
Self Test has a specificity of 99.8% in a performance evaluation conducted by untrained lay users.
This means a positive result will be correct 998 out of every 1000 tests.
A specificity study was performed on 1408 low or unknown risk and high risk individuals. Of the 1386
individuals identified as HIV negative using an approved comparator assay, 1376 were INSTI negative,
and 4 were invalid. The overall specificity of the INSTI HIV Self Test in fingerstick whole blood
specimens from the combined high risk and low or unknown risk populations, minus the invalid results,
was calculated to be 1376/1382 = 99.5% which means a positive result will be correct 998 out of every
1000 tests. (see Table 4)
Table 4- Performance of the INSTI HIV-1/HIV-2 Antibody Test on Fingerstick Whole Blood
Specimens from Individuals Presumed to be HIV Negative and Individuals at High Risk of HIV
Infection
Test Group
Total
specimens
INSTI
Non-
Reactive
INSTI
Reactive
INSTI
Invalid 1
Approved
Test Non-
Reactive
Approved
Test
Reactive
True
Negative 2
Low or
Unknown 626 620 6 0 626 0 626
Risk
High Risk 782 756 22 3 4 760 22 760
Total 1408 1376 28 4 1386 22 1386
1
Invalid results were not included in the calculation of specificity. The 4 specimens which gave
invalid results on INSTI were Non-Reactive on the approved test.
2
Reactives were confirmed by licensed HIV-1 Western Blot and excluded from the calculation of
specificity.
3 Of the 22 INSTI Reactive specimens, one was Non-Reactive by the approved test, i.e. INSTI false
Reactive.
Untrained Lay User Performance Evaluation
The performance of INSTI was evaluated in a prospective study conducted over 4 months at 3 different
sites. At each site, testing was conducted by non-professional lay users who had no laboratory
experience. The 11 people running the tests had no training on how to use the test. Fingerstick blood
from a total of 905 subjects with unknown HIV status and 483 subjects known to be HIV positive were
tested with INSTI and results compared to those determined by FDA approved reference methods.
The sensitivity of INSTI was 100% (517/517) and the specificity was 99.8% (869/871) There were no
invalid results reported (see Table 1).
Unrelated Medical Conditions and Potentially Interfering Substances
To assess the impact of unrelated medical conditions or potentially interfering substances on the
sensitivity and specificity of INSTI, 195 serum/plasma specimens from a cross section of medical
conditions unrelated to HIV-1 infection and 178 specimens with potentially interfering substances were
tested “unspiked” (HIV Nonreactive) and “spiked” with an HIV-1 positive specimen to give a low level
of reactivity in the INSTI HIV-1 Antibody Test. No cross-reactivity or interference with INSTI test
performance was observed with the following two exceptions:
1. Up to 5 specimens from individuals with multiple myeloma produced invalid INSTI results
depending on the INSTI kit lot tested.
2. Of the 20 specimens from individuals with elevated hemoglobin, one tested false Non-Reactive in
2 out of 3 INSTI kit lots.
Reproducibility Studies
The reproducibility of the INSTI test and ability of operators to consistently correctly interpret test
results was evaluated at 3 laboratory sites using 3 lots of INSTI on 3 separate days with 9 operators
(3 per site). A panel of 5 specimens, consisting of 4 HIV-1 antibody positive (one strong positive and
three low positives) and 1 HIV-1 antibody negative specimen was tested at each site. A total of 405
tests were conducted, 135 at each site, with a total of 81 tests per panel specimen. Overall all operators
interpreted the test results for each sample correctly, generating a reproducibility of the INSTI HIV Test
of 100% (405/405 samples tested).
FREQUENTLY ASKED QUESTIONS
What is HIV and AIDS?
HIV stands for Human Immunodeficiency Virus. HIV is the virus that causes AIDS (Acquired
Immunodeficiency Syndrome) if left untreated. When a person becomes infected with HIV, the virus
begins to attack his or her immune system, which is the body’s defense against illness. As a result,
that person becomes more susceptible to disease and infection.
When his or her body loses the ability to fight diseases, that person is diagnosed with AIDS. There is
no cure for HIV infection. However, treatment for HIV is highly effective.
How does someone get infected with HIV?
HIV spreads through contact with blood, semen, pre-seminal fluid, rectal fluid, vaginal fluids, or breast
milk of an infected person. Transmission can occur from unsafe sex. It can also result from exposure
to blood through the sharing of used syringes or needles. Women living with HIV can pass the virus to
their babies during pregnancy, childbirth, and breastfeeding. It is also possible to become infected with
HIV through a blood transfusion, although this is now very rare.
HIV cannot be passed on from one person to another through casual contact. There is no risk of
infection when we share everyday items such as food, dishes, utensils, clothes, beds and toilets with
a person living with HIV. The virus is not spread from contact with sweat, tears, saliva, or a casual kiss
from an infected person. People do not become infected from eating food prepared by a person living
with HIV. People have not become infected with HIV through insect bites.
How do I make sure I get enough blood?
Relax and have a drink of water about 20 minutes before you start the test. Warm your hands by
washing them with warm water. Ensure your hands are dry. Place your hand below waist level to
promote blood flow. Before using the lancet, look for a spot on the side of your finger tip that is smooth
and not calloused and away from your fingernail.
What is an antibody?
Antibodies are proteins produced by your body’s immune system in response to harmful organisms
like viruses and bacteria. Their purpose is to defend the body against infection by these organisms.
How accurate is the test?
Extensive research studies have shown that this test is extremely accurate when performed correctly.
In a recent study performed by untrained users, the test sensitivity was 100%.
It also has a proven specificity (a measure of reliability that the test will be negative for people who do
not have HIV infection) of 99.5%. In the untrained user study, the specificity was 99.8%.
*If you are unsure of your result you must go to a doctor to perform more testing.
The contents of Bottle 1, Bottle 2 or Bottle 3 do not absorb into the test device.
It is very rare for this to happen, but if it does, you will not be able to complete the test procedure and
read the results. You will need to perform another test.
What happens if I spill some of the contents of Bottle 1, Bottle 2 or Bottle 3 outside the test
device?
Keep going with the test procedure. As long as the control dot shows a visible dot after pouring Bottle
3 into the test device, the test results are valid.
How early can this test detect HIV?
Based on bioLytical's studies, INSTI demonstrates third generation performance and detects HIV
antibodies of the IgM and IgG class. IgM antibodies are the earliest antibodies that the body
produces after an HIV infection and are detectable within 21-22 days. 1,2 Depending on how quickly a
person’s immune system generates HIV antibodies after infection, it could still take up to 3 months to
get a positive result.
If you think you have been exposed to HIV within the last 3 months, and your results are negative,
you will need to test again after at least 3 months have passed since your exposure. The time from
HIV infection to when a test can correctly give a positive result is referred to as the ‘window period’.
1 Moshgabadi N, Galli RA, Daly AC, Ko SM, Westgard TE, Bulpitt AF, Shackleton CR., 2015.
“Sensitivity of a rapid point of care assay for early HIV antibody detection is enhanced by its ability to
detect HIV gp41 IgM antibodies.” J Clin Virol. 2015 Oct; 71:67-72.
2 M. Cohen, C. Gay, M. Busch, F. Hecht, The detection of acute HIV infection, J.Infect. Dis. 202 (2010)
270–277
I can’t see any dots.
Make sure you have adequate lighting. If no dots are visible, you may not have completed the test
correctly, or collected enough blood. You will need to perform another test.
How will I know if my test was done correctly?
The INSTI HIV Self Test has a built-in control dot to show that the test has been performed correctly
and that you have added the proper sample type and amount of blood sample. If the control dot does
not appear (invalid test result), your test has not worked. It is not possible to draw conclusions from
this result and you will need to perform another test. In the event of repeated invalid results, consult a
doctor.
M Manufactured by:
bioLytical TM Laboratories, Inc.
406 - 13251 Delf Place
Richmond, BC
Canada V6V 2A2
Phone: +1 604-204-6784
Fax: +1 604-244-8399
www.biolytical.com
Authorized Representative:
P
EMERGO EUROPE
Prinsessegracht 20
2514 AP, The Hague
The Netherlands
Phone: +31.70.345.8570
Fax: +31.70.346.7299
51-1200D 25-Sep-2020
© Copyright 2020, All rights reserved.
P.2
32

TRAINING PACK
BLOOD
COLLECTION
KIT
MULTIPLEX HIV-1/HIV-2
Syphilis Ab Test Kit
Product code: 8400
NHS Supply Chain Code: HHH2286
HIV-1/HIV-2
Antibody Test Kit
Product code: 8300
NHS Supply Chain Code: HHH208
www.hivhometest.com
www.biolytical.com
Designer: Josh
Job Name & Part Code: INSTI Training Manual Cov_ SC1374A v3
Date: 01/08/2019
Proof No.: #2
Dims: 297x210mm
Stock: White Card
GSM: 350
Finish: Matte
Special Finish(s):
300c
300c
300c
300c

INSTI ® HIV-1/HIV-2 Antibody Tests and Multiplex
HIV-1/HIV-2 Syphilis Ab Test are intended for use
by trained personnel in:
• Medical facilities
•
Doctors’ surgeries
• Clinical laboratories
•
Emergency care situations
• Voluntary counselling and testing programs
Now available for home use INSTI ® Self Test.
More information on this product inside.
USEFUL CONTACTS
Amber Matthews
Business Development Manager
South of England & Wales
T: 07967 105387
E: amber.matthews@pasantehealthcare.com
Tricia Frame
Business Development Manager
North of England, Scotland & N. Ireland
T: 07967 105388
E: tricia.frame@pasantehealthcare.com
Manufactured by:
bioLytical Laboratories
www.biolytical.com
Distributed by:
Pasante Healthcare Ltd,
Commerce Way, Lancing, West Sussex, BN15 8TA, UK.
T: +44 (0)1903 753844 www.pasantehealthcare.com
SC1374A v5