Lab Experiment 4a and 4b Flashcards
Initial lesion of Syphilis
Chancre
Syphilis is a venereal disease caused by a spirochete, the ______________.
Treponema pallidum
Two types of Antibodies
Treponemal and Non-treponemal antibody
Antibody specific for Treponema pallidum
Treponemal antibody
Antibody-like substance formed as a result of the interaction of the treponema with the body tissues
Non-treponemal antibody
Two major groups of test for Syphilis
Non-treponemal or Non-specific Reagin tests
And
Specific or Treponemal test
Discuss the principle of RPR CARD Test
RPR antigen suspension is a carbon particle cardiolipin antigen which detects “reagin”, an antibody-like subtance present in sera or plasma from syphilitic persons, and occasionally in persons with other acute and chronic conditions
What happens when a specimen contains antibody?
Flocculation occurs with a coaggulation of the carbon particles of the RPR antigen
How does flocculation looks like in RPR Test?
They appear as black clumps against the white background of the test card. Result can be read macroscopically.
Non-reactive specimens appear as what?
An even light-gray color
Plasma or serum samples must be free from?
Hemolysis and bacterial contamination
If the plasma or serum samples are not used immediately, they must be stored at what temperature?
2 - 8 C
If the plasma or serum samples are to be stored more than 24 hours, at what temperature must they be stored?
-20 C
Discuss the procedure of RPR CARD Test
Look at manual
Discuss the procedure of T. pallidum Hemagglutination Test (4B)
Look at manual
What is reagin?
An antibody-like substance present in the plasma or sera from syphilitic persons and occasionally from persons with acute and chronic conditions
How many minutes do we rotate the card on the mechanical rotator? And at what rpm?
8 minutes at 100 rpm.
How much does one drop of the dispenstir device deliver?
Approximately 0.05 ml/drop
What are the titles of Experiments 4a and 4b?
Rapid Plasma Reagin CARD Test (Qualitative Procedure) and Treponema pallidum Hemagglutination Test
In Expt 4b, how many uL of diluent must be placed in wells 1, 2 and 3?
10, 25 and 25 uL respectively
In Expt 4b, explain number 2 in the procedure.
Add 25 uL of serum sample to well 1. Mix then transfer 25 uL to well 2. Mix then transfer 25 uL to well 3. Mix then discard 25 uL from well 3.
In Expt 4b, how many uL of the well resuspended control cells are to be added to wells 2 and 3?
75 uL each
In Expt 4b, how many minutes do we leave the microtiter plates before reading the results?
45 - 60 minutes
