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Information
PRINCIPLE
The RPR Syphilis screening test is a macroscopic non-treponemal
flocculation card test for detection and quantify reagin, an antibody
like substrate present in serum or plasma from syphilitic persons.
SPECIMEN COLLECTION
Plasma unheated or heated serum may be used. Avoid hemolysis.
Serum samples are reportedly stable for 5 days if stored at +4°C.
plasma collected with EDTA is reportedly stable up to 24 hours.
REAGENT AND MATERIAL SUPPLIED
R1 : Carbon Antigen Suspension
R2 : Positive Control Serum
R3 : Negative Control serum
ACCESSORIES
20G Dispensing Needle (16µl/ drop)
Disposable Test cards
Disposable stirring rods
Disposable sample droppers
PROCEDURE : QUALITATIVE TEST
1. Bring all reagents and samples to room temperature.
2. Using the disposable sample dropper, dispense one drop
of serum or plasma onto a separate circle on the test card. Use
a fresh disposable sample dropper for each sample. Repeat step 2
using the positive and negative control sera.
3. Using the disposable stirring rod, spread the sample over
the entire area of the test circle.
4. Mix the carbon antigen well and place one drop of "free
fall" Antigen suspension onto each test specimen using 20G
Dispensing Needle.
5. Place the card on a rotator and rotate for 8 minutes at
100 rpm. Immediately after 8 minutes rotation, read the results
macroscopically in good light.
QUALITATIVE TEST RESULTS
REACTIVE: The presence of large aggregates in the center or
the periphery of the test circle.
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WEAKLY REACTIVE : The presence of small
or fine aggregates.
NON-REACTIVE : Smooth grey appearance with no aggregates
visible.
PROCEDURE:
QUANTITATIVE TEST
FOR EACH SPECIMEN TO BE TESTED :
1. Place 0.05 ml (50 µl) of 0.9% saline with a
pipette into test circles, numbered 2 to 5. DO NOT SPREAD SALINE.
2. Place 0.05 ml (50 µl) of specimen onto test circle
1.
3. Place 0.05 ml (50 µl) of specimen onto the test
circle 2. Prepare serial two-folds dilutions by drawing the mixture
up and down the pipette 5-6 times (avoid any bubble formation).
Transfer 0.05 ml from circle 2 to 3, to 4, to 5. Dispose 0.05 ml
from circle 5 after mixing.
4. Using a new stirring rod for each specimen, start at highest
dilution of serum (circle 5) and spread over entire area of test
circle. Proceed to circles 4,3,2 and 1.
5. Follow steps 4 to 5 in the Procedure of qualitative test.
If the last dilution (circle 5) is reactive, the dilution series
should be extended as follows.
1. Prepare a 1:50 dilution of non-reactive serum with on
0.9% saline. This is to be used for making 1:32 and higher dilutions
of specimens to be quantitated. Dispense 50 µl onto circles
numbered 7 to 10.
2. Prepare a 1:16 dilution of test specimen by adding 100
µl serum to 1500 µl of 0.9% saline. Mix thoroughly.
Dispense 0.05 µl of 1:16 dilution of test specimens onto circles
6 and 7.
3. On circle 7, mix the sample and diluent using the disposable
dropper by drawing the mixture up and down 5 to 6 times (avoid any
bubble formation). Transfer 50 µl of the mixed sample to the
next circle. Repeat the mixing procedure. Continue this serial dilution
to circle.
4. Dispose 50 µl from circle 10 after mixing.
REFRENCES
Hunter, E.F., W.E. Deacon and P.E. Meyer, 1964. an approved
FTA Test for Syphilis, the absorbtion Procedure (FTA-ABS).
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