Antigen Delivery System: 3ml Dropping bottle and Needle.

RPR Sufficient Test Cards and Disposable Pipette stirrers.

Latex Test Kit Materials required but not provided
Mechanical rotator set at 100 +/- 2 r.p.m. with humidity covered timing device,
automatic pipettes, test tubes, gloves, and a light source.
500100-100-31
500100-500-31 ASSAY PROCEDURE
NOTE: All specimens, control serum samples and reagent should be at
23-29 °C before use.
Qualitative Card Test
2-8°C 100 Tests or 500 Tests 1. The person performing this test should refer to the RESULTS section to
become familiar with the expected results before performing test.
Otherwise, perform test with the controls supplied to become familiar with
the expected results. Dispense 1 drop of EACH control onto separate
INTENDED USE circles of the test card and follow STEPS 3 to 5 below.
2. Dispense one drop of serum or plasma sample onto a separate circle on the
The Diagnostic Automation, Inc. (DAI) Rapid Plasma Reagin (RPR) Test is a test card with the disposable stirrer pipettes supplied. Use a fresh stirrer
non-Treponemal Flocculation Test that is used to detect and quantify reagin, an pipette for each sample. When using the stirrer pipette, hold it in a vertical
antibody present in serum or plasma from persons with syphilis, or with other position to ensure accurate delivery.
treponemal diseases. Occasionally individuals with other diseases or conditions 3. Using the flat end of the stirrer pipettes, spread the sample over the entire
may also be reactive in the non-Treponemal Tests. area of the test circle.
4. Mix the carbon antigen reagent well. Attach needle to the dropping bottle.
Squeeze the dropping bottle to release air and draw sufficient reagent into
the bottle. Discard the first few drops and then dispense 1 drop of the
SUMMAY & TEST PRINCIPLE antigen (while holding the bottle in a vertical position) to a test circle
Treponema pallidum, the etiologic agent responsible for syphilis produces at containing the sample. DO NOT MIX the sample and the antigen.
least two kinds of antibodies in human infections. Treponemal antibodies can 5. Place the card on an automatic rotator and place a humidity cover over
be detected by tests such as the Fluorescent Treponemal Antibody-Absorption card. Rotate at 100 r.p.m. for 8 minutes. Following rotation, a brief hand
(FTA-ABS) Test1 or MHA-TP whereas the reagin antibody is detected by non- rotation and tilting of the card (3 to 4 times) should be made to aid in
treponemal tests such as the RPR Antigen Card Test2. In the presence of the differentiating non-reactive from minimally reactive results. Read results
reagin antibody in the reactive sample, the RPR Antigen preparation will macroscopically in the “wet” state under a high intensity incandescent
produce flocculation consisting of black clumps against the white background lamp.
of the test card. By contrast, non-reactive samples will yield an even light-grey
homogenous suspension.
RESULTS
Qualitative
SPECIMEN COLLECTION AND PREPARATION Positive (Reactive) Result: A reactive result is indicated by the presence of
EDTA Plasma and unheated or heated serum may be used. Specimen should large aggregates in the centre or periphery of the test circle. All specimens
be free of bacterial contamination and hemolysis. Fresh, uncontaminated showing any degree of reactivity or roughness should be quantitated (follow
serum samples may be stored at 2-8 degree Celsius up to 5 days prior to Quantitative Test Procedure). Roughness is sometimes an indication of a
testing. Otherwise, the serum samples should be kept frozen. Plasma sample with a prozone.
specimens should be tested within 48 hours3, after that time the specimen Negative (Non-Reactive) Result: A non-reactive result will display a smooth
should be discarded. grey appearance.

MATERIALS AND COMPONENTS

Reactive Non-Reactive
Materials provided with the test kits Quantitative Card Test
RPR Carbon Antigen: *Suspension of Carbon containing 1. Dispense 1 drop (0.05 ml) of specimen using stirrer pipette onto circle 1.
approximately 0.003% Cardiolipin, 0.020% 2. Using an automatic 0.05 ml pipette (or stirrer pipette), dispense 1 drop of
Lecithin, 0.09% Cholesterol and 0.2g/L 0.9% saline onto circles to be numbered 2 to 5. DO NOT SPREAD.
Charcoal in buffer and less than 0.1% 3. Using an accurate volumetric pipette, dispense 0.05 ml of the test sample
sodium azide as preservative. onto circle 2. Insert the tip of the pipette into the resulting mixture and mix
Reactive Control: Human serum containing antibodies against them by drawing the mixture up and down the pipette approximately 8
Treponema Pallidium and less than 0.1% times. Avoid any bubble formation and transfer 0.05 ml of the mixed
sodium azide as preservative. sample to the third circle. Repeat this serial dilution procedure to circle 5
Non-Reactive Control: Human serum free of antibodies against and discard 0.05 ml from the last circle. Circles 1 to 5 now represent a
Treponema Pallidium and containing less dilution series as follows:
than 0.1% sodium azide as preservative.

COR CODE #31 Page 1 of 3

 Circle 1 2 3 4 5 quantitative tests were performed. No specific deviation trend has been
Dilution 1:1 1:2 1:4 1:8 1:16 observed.

Specimens DAI Test CDC Antigen Competitor Test

4. Using the flat end of the stirrer pipette, spread the diluted samples over the Positive 96 96 96
entire area of the test circles starting at circle no. 5 (highest dilution). Negative 109 109 109
Repeat this spreading procedure to circles 4,3,2, and 1.
5. Dispense 1 drop of carbon antigen from the dropping bottle to each circle. LIMITATIONS OF PROCEDURE
DO NOT MIX. Place the card onto the automatic rotator and rotate for 8
minutes. 1. The results of this test SHOULD NOT be used as a single diagnostic tool to
6. Immediately after 8 minutes of rotation, read results macroscopically in the make a clinical diagnosis. Instead, the test results must be evaluated
“wet” state under a high intensity incandescent lamp. The titre of the together with other clinical findings and observed symptoms to aid in the
sample is the reciprocal of the highest dilution to show macroscopic final diagnosis.
aggregates (see diagram in the RESULTS section). 2. False positive reactions occur occasionally with the RPR Carbon Antigen
7. If the sample is positive in the 1:16 dilution, the dilution series should Test. Such reactions sometimes occur in drug abuse and in such diseases
be extended as follows: as lupus erythematosus, mononucleosis, leprosy, viral pneumonia and after
a. Prepare a 1:50 dilution of non-reactive serum in 0.9% saline. smallpox vaccinations.
This is used to for making 1:32 and higher dilutions of 3. Reactive RPR Test specimens should be subjected to further confirmation
specimens to be tested. Dispense 0.05 ml of this diluent solution tests as recommended in the Manual of tests for Syphilis.
onto circles numbered 2 to 5. 4. Temperature of the reagents and specimens is critical to test outcome.
b. Prepare a 1:16 dilution of test specimen by adding 0.1 ml of 5. Plasma specimens over 48 hours old may give erroneous results.
serum to 1.5 ml of 0.9% saline. Mix thoroughly. Dispense 0.05
ml of 1:16 dilution of test specimen onto circles 1 and 2.
c. On circle 2, insert the tip of an automatic 0.05 ml pipette into the
resulting mixture (sample and diluent) and mix by drawing the PRECAUTIONS
mixture up and down the pipette approximately 8 times. Avoid
This product is for In Vitro Diagnostic Use Only. The bottle dispenser should
any bubble formation. Transfer 0.05 ml of the mixed sample to
be thoroughly washed and the needle should be rinsed with distilled water and
the next circle. Repeat the mixing procedure. Continue this serial
air dried after use. Each donor unit used in the preparation of this product has
dilution to circle no. 5 and discard 0.05 ml from this last circle.
been tested by an FDA approved method and found non-reactive for the
Circles 1 to 5 now represent a dilution series as follows:
presence of HbsAg and antibody to HIV Virus. Because no known test method
Circle 1 2 3 4 5 can offer complete assurance that hepatitis B virus, HIV Virus, or other
Dilution 1:16 1:32 1:64 1:128 1:256 infectious agents are absent, all human serum products and patient specimens
should be handled in accordance with good laboratory practices. The
d. Proceed with the test procedure described under STEPS 4 and preservative sodium azide may react with metal plumbing to form explosive
5 of the Quantitative Card Test. metal oxides. In disposal, flush with a large volume of water to prevent metal
e. Continue dilutions until an end-point titre is reached. azide build up.

RESULTS
1:2 1:4 1:8 1:16 1:32
STORAGE
When not in use, store reagents and controls at 2-8 degree Celsius. DO NOT
FREEZE. Prior to use, allow reagents and controls to warm up to 23-29 degree
Reactive Reactive Minimal Reactive Negative Negative Celsius. The antigen should be agitated gently to ensure homogeneity before
use. Remove only enough antigen from the bottle for the day’s testing use.
Expiration date is specified on the kit label and on each vial. Biological
indication of product instability is evidenced by inappropriate reaction of the
QUALITY CONTROL PROCEDURE carbon antigen reagent with the corresponding reactive and non-reactive
control sera.
The reactive and non-reactive controls have been included with the test kit to
monitor the performance of the reagent. If the expected results have not been
observed, the reagent should not be used.
REFERENCES
1. Hunter, E.F. Deacon, W.E. and Meyer, P.E., An improved FTA Test for
PERFORMANCE CHARACTERISTICS Syphilis, The Absorption Procedure (FTA-ABS). Public Health Reports,
79, 410-412, 1964.
The RPR Antigen has been tested according to Centers For Disease Control
2. Manual of Tests for Syphilis, Public Health Service Publication, No. 411,
(CDC) RPR Card Test Procedure. The RPR test was compared to the CDC
1969.
RPR Antigen and to another commercially available RPR Antigen in a clinical
3. Manual of Tests for Syphilis, Public Health Service Publication, No. 411,
evaluation. A total of 205 specimens were used in this study and the overall
1990.
agreement of the results was 100%. All 109 negative specimens gave negative
results when tested by all 3 types of RPR Antigen. The 96 positive specimens
gave positive results when tested by all 3 types of RPR Antigen. Only 4 out of
the 96 Positive specimens differed by 1 dilution difference in titre when

COR CODE #31 Page 2 of 3

 ISO 13485
ISO 9001

Diagnostic Automation/
Cortez Diagnostics, Inc.
21250 Califa Street, Suite 102 and 116,
Woodland Hills, California 91367 USA

Date Adopted 2016-01-22

500100-100-31 CORTEZ- RPR
500100-500-31 Latex Test Kit-2016
CEpartner4U, Esdoornlaan 13,
3951DB Maarn. The Netherlands.
www.cepartner4u.eu
Revision Date: 2009-09-01

COR CODE #31 Page 3 of 3