RPR-Carbon-DAC

Test for syphilis. AGGLUTINATION.

RPR-CARBON-CARDIOLYPINE ANTIGEN.

PRINCIPLE OF THE METHOD

The method is based on a precipitation reaction between a stabilized suspension of coal particles treated with a lipid complex and a number of antibodies present in serum or plasma of syphilis patients that, as a result of agglutination, form an antigen-antibody complex in the form of a precipitate (bunch) of the observed macroscopically.

The test is used in 2 variants: for rapid detection of syphilis (qualitative test) and for determining its content (semi-quantitative test).

SAMPLES FOR RESEARCH

Serum or plasma. Do not use hemolyzed and lipemic serums.

Stable at 2-8 ° C for up to 7 days, at minus 20 ° C - 3 months.

OPTIONAL EQUIPMENT

The dispenser is 10 - 50 μl. Rotator, frequency up to 100 rpm.

PRECAUTIONARY MEASURES

The control sera supplied in the kit were tested for antibodies to HIV, HCV and HBs antigen and were recognized as negative.

Possible reagent residues and serum samples of patients are to be destroyed in accordance with the approved nosocomial rules.

PROGRESS OF DEFINITION

Bring all reagents to 18-220C (room temperature), gently shake the bottle with RPR-Reagent until a homogeneous suspension is obtained and degrease the working surface of the slide.

Qualitative test (screening)

Mikroavoriant:

1. Place the 25 ml sample and 25 μl RPR-Positive Control and RPR-Negative Control in separate circles on the slide. (Note 1).

2. Add 10 μl RPR-Reagent to each circle without touching the sample and controls.

3. Mix the reagents thoroughly with a stick. For each sample use a new wand.

4. Rotate the slide for 8 minutes in regular circular motions so that the mixture rotates slowly inside the circle.

5. After 8 minutes, evaluate the result of the reaction under bright artificial light.

To standardize the rotation procedure, it is recommended to use a rotator (80-100 rpm).

If necessary, the volume of reagents and samples can be proportionally increased to 20-50 μl.

Macrovariate:

1. Place 50 ml of the sample in a circle on the slide.

2. Add 20 μl RPR-Reagent to each circle, without touching the sample and controls.

Then proceed the same way as the micro version.

For more convenient dosing of the reagent, use a dropper bottle with RPR-Reagent, in this version the volume of the sample (control) should be increased to 125 ml.

Semi-quantitative test

(definition of the title)

Prepare dilutions of the test serum in physiological saline solution 9 g / l. Then proceed similar to the qualitative test.

EVALUATION OF THE RESULTS

Explore visually the presence or absence of clots in circles, within a minute of extraction of the slide from the rotator. The results are recorded and recorded according to the following criteria:

Observed agglutination

Result

Medium or large clots

Positive

Small clumps

Weakly positive

No clots or light turbidity

Negative

Positive serum can be titered. For titration, prepare serial double dilutions with saline solution. The serum titer is defined as the largest dilution yielding a positive result.

QUALITY CONTROL

It is recommended that RPR-Reagent be monitored regularly with positive and negative control sera.

CHARACTERISTICS OF THE PROCEDURE

- The effect of the prozone is not observed until the titer ≥ 1/128.

-Sensitivity: 86% (primary syphilis) and 100% (secondary syphilis).

- Specificity: 98%.

Interference:

Hemoglobin up to 10 g / l, bilirubin up to 20 mg / dl and lipids up to 10 g / l do not affect the result of the determination. Some drugs and substances can influence the result of the determination4.

CLINICAL VALUE AND LIMITATIONS OF THE METHOD

Syphilis is a sexually transmitted disease caused by pale treponema (Treponema Pallidum), which occurs with periodic remissions and exacerbations, as well as the formation of foci of inflammation in tissues and organs.

Infection with pale treponema can lead to:

- to complicated balanitis, balanoposthitis, phimosis in men in the primary period;

- In the secondary period, syphilitic hair loss is possible, as well as damage to bones, periosteum and joints;

- Tertiary period is characterized by irreversible lesions of internal organs and tissues.

Cardiolipin-like tests are not specific for syphilis. All reactive samples should undergo additional serological tests (EIA, TPHA), to confirm the results.

False positive reactions can be detected with infectious mononucleosis, pneumonia, toxoplasmosis, pregnancy and autoimmune diseases.

The test is especially useful for determining the effectiveness of antibiotic therapy.

Delivery time: 5 days

The information is relevant to: 03.02.2018