ABBOTT PRISM® HTLV-I/HTLV-II - ILEX Medical Systems

RESULTS
Calculation of Cutoff and S/CO Values
The ABBOTT PRISM System calculates the ABBOTT PRISM HTLV-I/HTLV-II
assay cutoff value using the following formula:
Cutoff = Mean Negative Calibrator Net Counts
+ (0.15 x Mean Positive Calibrator Net Counts)*
Example: If the Mean NCC = 1,100, and the Mean PCC = 6,900,
1,100 + (0.15 x 6,900) = 2,135
Cutoff = 2,135
* The Positive Calibrator, HTLV-II Positive Assay Control (1), and Negative
Calibrator Mean Net Counts are calculated using the two lowest replicates. An
Instrument Code (02-211) will be displayed in place of the count value for the
third replicate. Each of the two remaining replicates of the Positive Calibrator
and the Negative Calibrator used to calculate the cutoff must meet all
specifications.
The ABBOTT PRISM System calculates the ABBOTT PRISM HTLV-I/HTLV-II assay
S/CO using the following formula:
S/CO = Sample Net Counts ÷ Cutoff
Example: If the Sample Net Counts = 3,000, and the Cutoff = 2,135,
3,000 ÷ 2,135 = 1.41
S/CO = 1.41
INTERPRETATION OF RESULTS
In the ABBOTT PRISM HTLV-I/HTLV-II assay, specimens with Net Counts less
than the cutoff value are considered nonreactive and need not be tested further.
Specimens with Net Counts greater than or equal to the cutoff value are considered
initially reactive. All specimens that are reactive on initial testing must be centrifuged
according to the table in the Specimen Collection and Preparation for Analysis
section of this package insert and retested in duplicate.
NOTE: If re-testing a specimen within 24 hours of the initial centrifugation, the
specimen is not required to be re-centrifuged.
If repeat testing shows the Net Counts for both retests to be less than the cutoff
value, the specimen is nonreactive. If the Net Counts of either retest are greater
than or equal to the cutoff value, the specimen is repeatedly reactive. Repeatedly
reactive specimens should be investigated further by supplemental tests such as
immunoblots, immunoprecipitation or immunofluorescent assays.
ABBOTT PRISM reports display sample results in Net Counts and/or S/CO. Net
Counts are used by ABBOTT PRISM to interpret results. The S/CO value is provided
in reports to show relative reactivity to the cutoff value. In the ABBOTT PRISM
HTLV-I/HTLV-II assay, specimens with S/CO values of less than 1.00 are considered
nonreactive. Specimens with an S/CO value of greater than or equal to 1.00 are
considered reactive.
For details on configuring the ABBOTT PRISM System to use Grayzone
Interpretations, refer to the ABBOTT PRISM Operations Manual, Section 2.
READING RESULTS
Some S/CO values may be flagged with "" symbols. Refer to the ABBOTT
PRISM Operations Manual, Section 5.
SYSTEM ERRORS
For a description of the error codes that appear in the ABBOTT PRISM Report,
refer to the ABBOTT PRISM Operations Manual, Section 10.
LIMITATIONS OF THE PROCEDURE
• Testing of previously frozen samples with the ABBOTT PRISM HTLV-I/HTLV-II
assay may cause an increase in non-specific reactivity.
• The ABBOTT PRISM HTLV-I/HTLV-II assay does not discriminate between
HTLV-I and HTLV-II antibody reactivity.
• This assay was designed and validated for use with human serum or plasma
from individual patient and donor specimens. Pooled specimens must not be
used since the accuracy of their test results has not been validated.
• It is recommended that repeatedly reactive specimens be investigated by
supplemental testing. Individuals who are repeatedly reactive should be referred
for medical evaluation which may include additional testing.
• False reactive results can be expected with any test kit. Falsely elevated results
have been observed due to non-specific interactions (see Table II).
• Performance has not been established for body fluids other than serum or
plasma.
• Previously frozen specimens must be centrifuged per the Specimen Collection
and Preparation for Analysis section of this package insert prior to running the
assay.
• Serum from heparinized patients may be incompletely coagulated. Erroneous
or inconsistent results may occur due to the presence of fibrin. To prevent this
phenomenon, draw specimen prior to heparin therapy.
• The use of specimens with obvious microbial contamination should be avoided.
• Although the association of infectivity and the presence of anti-HTLV-I/HTLV-II
is strong, it is recognized that presently available methods for
anti-HTLV-I/HTLV-II detection are not sensitive enough to detect all potentially
infectious units of blood or possible cases of HTLV-I/HTLV-II infection.
6
• The interpretation of results of specimens found repeatedly reactive by the
ABBOTT PRISM HTLV-I/HTLV-II assay and negative or indeterminate on
additional supplemental testing is unclear. Further clarification may be obtained
by testing another specimen from the same patient taken 3 to 6 months later.
EXPECTED RESULTS
In a random population of 6,678 volunteer blood donor specimens, four (0.06%)
were reactive by the ABBOTT PRISM HTLV-I/HTLV-II assay. Among 148
preselected anti-HTLV-I positive and 148 preselected anti-HTLV-II positive
specimens, the ABBOTT PRISM HTLV-I/HTLV-II assay detected 100.00% as
repeatedly reactive.
SPECIFIC PERFORMANCE CHARACTERISTICS
Precision
Assay reproducibility was determined by assaying a 28 member panel consisting
of four replicates each of three diluted specimens reactive for anti-HTLV-I (panel
members 1, 2, and 3), three diluted specimens reactive for anti-HTLV-II (panel
members 4, 5, and 6) and one specimen nonreactive for anti-HTLV-I or anti-HTLV-II
(panel member 7). The panel was tested in five runs over five days with each of
three master lots at a total of three sites.
The intra- and inter-run standard deviation (SD) and percent coefficient of variation
(%CV) were analyzed with a variance components analysis, using a nested analysis
of variance model67 (Table I).
Mean S/CO is defined as the mean sample net counts (NET) divided by the
calculated Cutoff.
TABLE I
ABBOTT PRISM HTLV-I/HTLV-II
Assay Reproducibility
Number of Mean Intra-run Inter-run*
Panel Member Replicates S/CO SD %CV SD %CV
1 120 8.27 0.339 4.1 0.487 5.9
2 118 5.19 0.240 4.6 0.291 5.6
3 114 3.17 0.131 4.1 0.190 6.0
4 119 6.90 0.291 4.2 0.446 6.5
5 120 3.98 0.180 4.5 0.254 6.4
6 119 2.35 0.096 4.1 0.138 5.9
7 120 0.56 0.026 4.7 0.033 5.9
Negative Control 240 0.42 0.031 7.3 0.035 8.3
Positive Control 178 2.02 0.114 5.7 0.142 7.0
Supplemental
Positive Control
240 1.84 0.097 5.3 0.115 6.3
Number of Mean Intra-run Inter-run*
Calibrators Replicates NET SD %CV SD %CV
Negative 180 833 50.1 6.0 82.8 9.9
Positive 179 7,208 292.6 4.1 485.6 6.7
Supplemental
Positive
180 6,179 273.5 4.4 438.2 7.1
* This term includes intra-run variability.
Specificity
The specificity of the ABBOTT PRISM HTLV-I/HTLV-II assay was estimated
assuming a zero prevalence of HTLV-I and/or HTLV-II in volunteer blood donors
and plasmapheresis donors.
A total of 6,678 serum and plasma specimens from volunteer blood donors and
plasmapheresis donors was collected from four blood centers (Table II). Of the
four repeatedly reactive specimens one was excluded as confirmed positive by
DBL HTLV-I/II Western Blot, RIPA-I, and/or RIPA-II. a Therefore, of the 6,677
donations presumed seronegative for anti-HTLV-I and anti-HTLV-II, ABBOTT
PRISM HTLV-I/HTLV-II had an estimated specificity of 99.96% (6,674/6,677) with
a 95% confidence interval of 99.87% to 99.99%.
Specimens from individuals with medical conditions unrelated to HTLV-I and/or
HTLV-II infection or containing potentially interfering substances and specimens
from random hospital patients were tested with the ABBOTT PRISM HTLV-I/HTLV-II
assay (Table II).
a A confirmed positive result was defined by the presence of antibodies to two gene
products (gag p24 and env gp46 or gp61/67) using Western Blot and/or RIPA.