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Causative Agent
Treponema pallidum Slender, tightly coiled, pointed-end, 6-10 axial filamentous gram negative rod
Microaerophilic, obligate parasite Requires mammalian cells for survival Sheath cover aids its pathogenicity by reducing or limiting antigenic stimulation of the hosts immune system Does not survive in citrated blood at 4C for more than 72 hrs.
T. pallidum subspecies pallidum - syphilis T. pallidum subspecies pertenue - yaws T. pallidum subspecies endemicum - nonvenereal (endemic) syphilis/bejel T. carateum - pinta
Species- indistinguishable serologically and morphologically Differentiation- based on clinical & epidemiological characteristics
Diagnostic Evaluation
Fluorescent Treponema pallidum-absorbed (FTA-ABS) Microhemagglutination Treponema pallidum (MHA-TP) Passive Agglutination Treponema pallidum test (TP-PA)
Measles Chicken pox Hepatitis Infectious mononucleosis Leptospirosis Malaria Leprosy Rickettsial disease Trypanosomiasis Lymphogranuloma venereum (LGV)
Noninfectious conditions:
RPR test
The RPR test is a non treponemal testing procedure for the serologic detection of syphilis
Principle of RPR
Reagin
The reagin (Ag) binds to the test antigen which consists of cardiolipin-lecithin coated particles macroscopic flocculation
An antibody like substance present in serum or plasma from individuals with syphilis
Antibodies
Begin to appear in blood 4 to 6 weeks after infection IgG and IgM antibodies
Test Cards RPR Control Cards RPR Antigen Distilled Water Dispenstirs Rotator
plastic coated card that consist of several circles that are 18 mm in diameter Controls (strongly reactive, weakly reactive, and non-reactive) are contained on each kit
The test results should be reported as reactive (even if minimally reactive) or nonreactive
rotated manually by hand 3 to 4 rotations and then read immediately macroscopically in the wet state under a high intensity lamp
Explanation of Results
A negative RPR test may indicate one of the following: 1. The patient does not have syphilis 2. The infection is too recent for antibodies to be produced. (Repeated tests should be administered at 1 week, 1 month, and 3 month intervals to establish presence or absence of disease) 3. The syphilis is latent or inactive 4. Faulty immunodefense mechanism 5. Faulty lab techniques
Explanation of Results
A positive reaction is not conclusive for syphilis. Several conditions produce biologic false positive results for syphilis (False positive means that the test revealed a positive reaction when it was actually negative) False positives may reveal the presence of other serious diseases
positive RPR tests may be resolved by testing the patients serum with a specific treponemal antigen test
pallidum
TP-PA
Passive Particle Agglutination Test for Detection of antibodies to Treponema pallidum
Principle of TP-PA
The
kit is manufactured using gelatin particle carriers sensitized with purified pathogenic T. pallidum (Nichols strain) Sensitized particles are agglutinated by the presence of antibodies to T. pallidum in human serum / plasma
A. Reconstituting solution B. Sample Diluent ( liquid ) C. Sensitized particles ( lyophilized ) D. Positive control
( liquid )
E. Droppers (25ul)
1
100 25 1:5
2
25 25 1:10
3
25 25 1:20 25
4
25 25 1:40
DISCARD
Test Specimen Dilution Unsensitized Particle (ul) Sensitized Particle (ul) Final Dilution
25 1:40 1:80
INTERPRETATION OF RESULTS
SETTING PATTERNS OF PARTICLE READING INTERPRETATION
Particles concentrated in the shape of a button in the center of the well with a smooth round outer margin.
Particles concentrated in the shape of a compact ring with a smooth round outer margin. Definite large ring with a rough multiform outer margin and peripheral agglutination. Agglutinated particles spread out covering the bottom of the well uniformly.
( - )
NEGATIVE
( + / - )
INDETERMINATE
( + )
POSITIVE
( ++ )
NEGATIVE (-)
INDETERMINATE (-/+)
POSITIVE ( + / ++)
FINAL DILUTION
REACTION
1:40 1:80
(-) (+)
POSITIVE
FINAL DILUTION
REACTION
1:40 1:80
(-) (-)
NEGATIVE
ARCHITECT Syphilis TP
ARCHITECT
Syphilis TP assay
Chemiluminescent Microparticle Immunoassay (CMIA) for the qualitative detection of antibody to Treponema pallidum in human or plasma as an aid to diagnosis of syphilis.
REAGENTS:
1. Microparticles 2. Conjugate 3. Assay Diluent
4. Pre-trigger Solution
5. Trigger Solution 6. Wash Buffer