1. Remove the antigen suspension vial and one control card envelope from the refrigerator. Allow the items to warm to room temperature.

2. Resuspend the contents of the vial by vigorously shaking the antigen vial.

3. Snap the neck of the vial.
4. Attach the needle (provided in the kit) to a 1 ml tuberculin syringe. Slowly draw up into the syringe from the vial approximately 1 ml of the antigen suspension.

5. Hold the syringe perpendicular to the surface and count the number of drops dispensed from a 0.5 ml volume. Allow the drops to fall into the antigen vial. NOTE: The needle is accurate if 30 drops, plus or minus 1 drop, are dispensed from the 0.5 ml volume.

6. Slowly expel the remainder of the antigen solution in the syringe back into the antigen vial.

7. Remove the needle from the syringe. Place the needle on the tapered fitting of the plastic dispensing bottle (provided in the kit).

8. Slowly withdraw all the contents of the antigen vial by collapsing the dispensing bottle and using it as a suction device.

9. Allow the rotator to warm up for 5 to 10 minutes; adjust to 100 rpm.

RPR Test Procedure
To detect syphilis using the RPR Card test, follow the steps below:

1. Open the foil package and remove the control card.

2. Use a pipette/stirrer to reconstitute each control card circle with 0.5 ml of distilled water.

3. Mix solution in control card circle with pipette/stirrer until the dehydrated control specimen is dissolved. Spread specimen over entire area of circle. Use a separate pipette/stirrer for each circle.

4. Draw the patient's sample by holding the pipette/stirrer between the thumb and forefinger near the stirring or sealed end and squeeze. Do not release pressure until the open end is below the surface of the specimen, then release pressure to draw up the sample.

5. Hold the pipette/stirrer in a vertical position, directly over the card test area where the specimen is to be delivered; squeeze pipette/stirrer, allowing 1 drop to fall onto the test area.

6. Invert the pipette/stirrer and, with the sealed end, spread specimen within the circle. Discard the pipette/stirrer when done.

7. Continue the steps above until one or two test cards are filled with patient's samples.

8. Gently shake the antigen dispensing bottle before use. Hold the bottle in the vertical position and dispense several drops into the dispensing bottle cap to ensure the needle passage is clear. Allow 1 "free-falling" drop to fall onto each test area. Do not stir; the mixing of antigen suspension and specimen is accomplished by rotation.

9. Put the card(s) on the rotator and cover with the humidifying cover. Rotate cards for 8 minutes at 100 rpm. To help differentiate nonreactive from reactive results, you should briefly rotate and tilt the card by hand (3 or 4 back-and-forth motions).

10. Immediately read the card macroscopically (with the unaided eye) in the "wet" state under a high-intensity lamp.

11. Compare the patient's tests to the controls for correct interpretations. The reactive control should show small to large clumps. The nonreactive control should show no clumping or very slight roughness. The reactive-minimal- to-moderate control should show slight but definite clumping.

12. Report the test as

reactive, if agglutination or flocculation is present, or

nonreactive, if no agglutination is present.
NOTE: The RPR Card test is used as a screen for syphilis. If a patient's RPR is reactive, the patient should be sent to a laboratory to have a FTA-ABS (Fluorescent Treponemal Antibody Absorption Test) performed. The FTA-ABS, a more precise test, is used to confirm primary, secondary, and late syphilis.