"Syphilis-AgKL-RMP" Cardiolipin antigen for microprecipitation reaction Set No. 3

APPOINTMENT

The kit is used in the diagnosis of syphilis to study the plasma (serum) of the blood or cerebrospinal fluid (CSF) of a person in a microscopic reaction (RMP).

PRINCIPLE OF THE METHOD

The test is based on the interaction of the cardiolipin antigen (AGCL), similar to the Treponema pallidum lipoprotein antigen, with the corresponding antibodies (reagins) that appear in plasma (serum) of untreated patients after 2-3 weeks, and in cerebrospinal fluid - after 4-8 weeks after infection.

The interaction of AGCL with reagins leads to microprecipitation reaction (loss of flakes of different size) and is recorded visually.

COMPOSITION OF THE KIT:

Set No. 3 (Cat. No. 03.07.4) includes:

AgCl suspension - AgCl suspension in 10% choline chloride solution containing cardiolipin - 0.033%; lecithin - 0.27%, cholesterol - 0.9%, EDTA (stabilizer) at a final concentration of 0.0125 mol / l and thimerosal (preservative) at a final concentration of 0.1%. Milky-white suspension, when settled, separates into an opalescent colorless liquid and a dense white precipitate

cat 03.07.4 - 1 bottle (10 ml each).

The basic version of the set number 3 is designed for the study of 500 samples (Cat. No. 03.07.4)

At the request of the consumer, the basic equipment of the kit (the number of packages with reagents and their volume) can be changed.

ANALYZABLE SAMPLES

The reaction is carried out with plasma (serum inactivated at 561 ° C for 30 minutes) of blood or CSF, obtained by standard methods.

Samples of plasma (serum) of blood with pronounced hemolysis, hyperlipidemia and bacterial growth are not subject to study.

Samples intended for research can be stored from 2 to 8 ° C for no more than 7 days, their long-term storage in a frozen state at a temperature of minus 20 ° C and below is permissible. Repeated freezing of thawed samples is not allowed.

PRECAUTIONS FOR WORKING WITH THE KIT

The kit is biologically safe, but test specimens should be treated as potentially infectious materials.

EQUIPMENT, MATERIALS AND REAGENTS:

· Laboratory centrifuge, providing an angular speed of 1000-2000 rpm;

· Laboratory shaker;

· A thermostat or a water bath providing the heating temperature (37 + 1) 0С or (56 + 1) 0С, respectively;

· Stopwatch;

· Pasteur pipettes;

· Pipettes, allowing to take liquid volumes of 0.02 and 5.0 ml;

· Test tubes with a capacity of 10 ml;

· Purified water (distilled or deionized);

· 0.9% solution of sodium chloride;

· Glass or plexiglas plate;

· Rubber or plastic gloves.

PREPARATION OF REAGENTS FOR ANALYSIS

Kit number 3

Before using the reagents for at least 30 minutes at a temperature of from 18 to 25 ° C, stir the AgCL suspension thoroughly until a homogeneous suspension is formed.

ANALYSIS CONDUCT

Qualitative method

Kit number 3

60 μl of the test sample is applied to ordinary glass or a hollow plate of plastic, then 20 μl of antigen emulsion is added. Place the glass or plate on the shaker platform and rotate in the horizontal plane for 8 minutes, then immediately take into account the results of the reaction (the optimum reaction temperature is 23-28 ° C).

Records of the results of the reaction

In the study of the sample from patients with syphilis, there is a positive reaction in the form of falling out flakes of various sizes, estimated at the crosses (large (++++) and medium (+++) with clear clarification of the liquid — positive reaction, small (++) - weakly positive ), and with plasma or inactivated serum from healthy individuals there is a negative reaction in the form of opalescence.

The results of the reaction are taken into account visually when lighting is not lower than 300 lux.

Documented recording of results is possible using the Expert-Lab hardware and software system.

Semi-quantitative method (determination of reagin titer)

The titer of reagins is determined only in the studied samples, which gave a positive or weakly positive reaction.

1. The test sample diluted with saline in the ratio of 1: 1, 1: 2; 1: 4, 1: 8, 1:16, 1:32.

2. Each dilution to explore in the same way as in the qualitative method.

The titer of reagins in the sample to be examined shall be the maximum dilution with which a positive or weakly positive reaction is obtained, subject to a negative reaction with subsequent dilution. If a positive reaction is obtained with the maximum dilution used, a series of dilutions must be continued to determine the titer of reagins.

SHELF LIFE. STORAGE AND TRANSPORTATION CONDITIONS

Kit number 3

Shelf life - 1.5 years.

The kit should be stored in the manufacturer's packaging at a temperature of 2 to 8 ° C for the entire shelf life. Freezing not allowed