3. Using the broad end of the dispenstir, mix until the dehydrated control specimen is dissolved. Spread specimen over entire area of circle. Use a separate dispenstir for each circle.
4. To draw the patients sample, hold the dispenstir between the thumb and forefinger near stirring or sealed end and squeeze; do not release pressure until the open end is below the surface of the specimen. Release finger pressure to draw up the sample.
5. Hold the dispenstir in a vertical position, directly over the card test area to which the specimen is to be delivered; squeeze dispenstir, allowing 1 drop to fall onto the test area.
6. Invert the dispenstir, and, with the sealed end, spread specimen within the confines of the circle. Discard the dispenstir.
7. Continue the above steps until one or two test cards are filled with patients samples.
8. Gently shake the antigen dispensing bottle before use. Hold in the vertical position and dispense several drops into the dispensing bottle cap to ensure that the needle passage is clear. Allow 1 free-falling drop to fall onto each test area. Do not stir; mixing of antigen suspension and specimen is accomplished during rotation.
9. Put card(s) on rotator and cover with humidifying cover.
10. Rotate for 8 minutes at 100 r.p.m. Following rotation, to help differentiate Nonreactive from Reactive results, a brief rotating and tilting of card by hand (3 to 4 to-and-fro motions) must be made.
11. Then immediately read Card microscopically in the wet state and under the high-intensity lamp.
12. The Reactive control should show characteristic strong clumping; the Nonreactive control should show smooth, grayish appearance of unclumped particles. The Reactive Minimal-to-Moderate control should show minimal-to-moderate clumping. The patients tests should be compared to the controls for correct interpretations.
13. Report test as:
a. Reactive if specimen shows agglutination or flocculation.
b. Nonreactive if specimen shows no agglutination at the end of 8 minutes rotation.
c. If the RPR test is reactive, an FTA-ABS (Flourescent Treponemal Antibody Absorption Test) must be run on the specimen.
The main reason for including this test is that mononucleosis imitates many diseases so well that diagnosis is confirmed only by selective serologic testing.
1. Absorption of serum with a suspension of a guinea pig or horse kidney antigen removes antisheep agglutinins in the serum of patients with serum diseases and various infectious diseases.
2. In some serum of patients with infectious mononucleosis, a substantial part of the antibodies remains after absorption.
3. Absorption with a suspension of beef cells removes the antisheep agglutinins in infectious mononucleosis, but leaves them in other infectious diseases.
Rapid slide tests for infectious mononucleosis are based on these principles. Suspensions of guinea pig kidney and beef erythrocyte stomata result in satisfactory instant absorption of antibodies and clear differentiation between infectious mononucleosis and noninfectious mononucleosis sera. Infectious mononucleosis antibodies may be demonstrated as early as the fourth day of illness and practically always by the twenty-first day. Positive results may continue for several months.
1. On a clean slide (supplied with kit) place 1 drop of guinea pig antigen, reagent I, into box number 1.
2. Place 1 drop of the beef erythrocyte stomata, reagent II, into box number 2.
3. Add 1 drop of test serum on plasma to both boxes. Mix each with separate sticks.
4. Add 1 drop of horse erythrocyte antigen (supplied with kit) to both boxes. Mix each with separate disposable sticks.
5. Rock slide back and forth for 2 minutes so that liquid flows slowly over the entire area of the boxes.
6. Read results after 2 minutes.
a. Agglutination in box 1 is positive for infectious mononucleosis.
b. Agglutination in box 2 is positive for noninfectious mononucleosis.
c. No agglutination in either box is negative for mononucleosis.