MIKROGEN recomBlot Treponema IgG recomBlot Treponema IgM ...

E
Western blot
Treponema pallidum
C
recomBlot Treponema IgG
recomBlot Treponema IgM
Immunoblot test with antigens produced by recombinant techniques for the detection
of IgG or IgM antibodies against Treponema pallidum.
Treponema pallidum subsp. pallidum is the causative agent of Syphilis, a chronic disease
occuring only in humans. The disease progresses through stages. After a short
propagation period at the site of infection (primary stage) the hematogenic spreading
of the pathogen throughout the entire organism follows (secondary stage). After the
decline of acute symptoms Treponema pallidum can persist for years or lifetime in the
human organism (stage of latency) and then can cause late complications (tertiary
Syphilis). During pregnancy the pathogen can be transmitted to the fetus (Syphilis
connata). In approximately 40 % of the cases infection results in abortion or death of
the child shortly after birth. The propability of infection in utero is around 40-70 %.
The serologic diagnosis of syphilis intends the use of a sensitive antibody screening
assay in the first step (TPHA, ELISA). In the case of equivocal or positive results in the
screening test a confirmation assay is performed in the second step to assure the
specificity (FTA-ABS, immunoblot). Here the immunoblot plays a very important role,
as only with the western blot technique the determination of antibody production against
essential Treponema pallidum antigens is possible. At the same time the detection of
different antibody classes is possible. With the use of recombinant antigens the
recomBlot Treponema IgG/IgM meets all these requirements perfectly.
MIKROGEN
molekularbiologische Entwicklungs-GmbH
React. Control
Tp 47
TmpA
Tp 37
Product Advantages
• Recombinant Antigens, therefore
➣ Highest sensitivity and specificity
➣ Simple and clear interpretation due to easy to read bands
➣ Use of pathogen specific Treponema antigens Tp 47, Tp17 and Tp 15 as well as TmpA
Easy test procedure; automation possible
Safe evaluation due to kit specific control strip as well as cut-off control
Separate detection of IgG and IgM antibodies possible
Complement as confirmation tests ideally the recomWell Treponema tests (ELISA)
C label: The recomBlot Treponema tests meet the high standard of the EC directive 98/79/EC on in vitro
diagnostic medical devices
Recombinant Antigens
Recombinant Antigen
F unction
Size
[ kDa]
Tp 47
Membrane protein
47
TmpA Membrane protein
42
Tp 37
Flagellin A
37
Tp 17
Membrane protein
17
Tp 15
Membrane protein
15
Tp 17
Tp 15

Test Principle and Procedure
E E E
Evaluation
Blood
donors
( TPHA
negative)
Potential
cross
reactive
sera
( Borreliosis,
leptospirosis,
auto
immune
dis.
, pregn.
women)
Specificity
recomBlot Treponema
IgG IgM
total n egative
equiv. positive total n egative
equiv. positive
205 202 3 0 205 205 0 0
51 50 1 0 49 47 2 0
C alculation
of
positive
findings
only
1 00
%
100
%
C alculation
of
equivocal
findings
as
positive
9 8,
4 %
99,
2 %
TPHA positive/
IgG
FTA-ABS
positive
84 0 3 81
recomBlot Treponema
IgG IgM
total n egative
equiv. positive total n egative
equiv. positive
Positive for
IgM
antibodies
( 19S-IgM-FTA)
46 0 7 39
Sensitivity
1 st. Incubation: A test strip loaded with Treponema antigens is incubated with
diluted serum or plasma in a dish for 1 h.
Wash 3 times
2 nd. Incubation: Peroxidase conjugated anti-human antibodies (IgG or IgM specific)
are added. Incubate for 45 min.
Wash 3 times
3 rd. Incubation: 5 - 10 minutes after addition of the coloring solution, insoluble
colored bands develop at the sites on the test strips occupied by
antibodies.
In collaboration with an european expert for Treponema pallidum, Prof. Dr. H.-J. Hagedorn, more than 300 serum probes
with defined TPHA status were analysed with recomBlot Treponema IgG/IgM. TPHA negative serum probes were used
for calculation of specificity and TPHA and IgG FTA-ABS positive serum probes were used for calculation of sensitivity.
C alculation
of
positive
findings
only
9 6,
4 %
84,
8 %
C alculation
of
equivocal
findings
as
positive
1 00
%
100
%
Storage and Shelf Life At 4 °C 18 months from the time of production
Commercial Product Article No. 5102 recomBlot Treponema IgG
Reagents for 20 determinations
Article No. 5103 recomBlot Treponema IgM
Reagents for 20 determinations
MIKROGEN GmbH
Floriansbogen 2-4 · D-82061 Neuried · Germany · Tel.: +49 (0)89 54801-0 · Fax: +49 (0)89 54801-100
Internet: www.mikrogen.de · eMail: mikrogen@mikrogen.de
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