—Plastic dispensing bottle —20-gauge, galvanized needle, blunt cut —Test cards —Pipette/stirrers, 50 microliter (Fl)  One 1 ml tuberculin syringe  Distilled water  Mechanical rotator (adjusted to 100 rpm)  RPR Card Test Control Cards (each consisting of three labeled test areas containing lyophilized (meaning a stabilized preparation of a biological substance, such as blood, that has been frozen rapidly and then dehydrated under a high vacuum) control specimens with designated patterns of reactivity: Reactive, Reactive- Minimal-to-Moderate, and Nonreactive.) NOTE: RPR Card Test Antigen and Control Cards must be stored at 4EC when not in use. Both items are stable until the expiration date. Store “in use” antigen suspension in the dispensing bottle at 4EC. The antigen suspension is stable for 3 months or until the expiration date, whichever occurs first. Preliminary Preparations for RPR Test The following preliminary preparations must be performed before RPR testing can begin: 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 7-30