SU1258416A1 - Method of determining markers of virus heratitis - Google Patents

Description

This invention relates to medicine and can be used to diagnose human viral hepatitis.

The purpose of the invention is to simplify the process while maintaining its high sensitivity by using a polyethylene film as a solid phase and detecting radioactivity by autoradiography.

The method is carried out as follows.

Polyethylene film (PEP) in the form of household bags of polyethylene household 400x230 cm, thickness 0.02 mm, price S Kop is suitable as the solid phase. Polyethylene film of any thickness is suitable,

PETs are lined with a ball pen with inter-perpendicular lines with an interval of 1 cm. Then the PETs are placed on a glass plate (with the lined side down) covered with wet filter paper. The points of intersection of the lines serve as the points of application of the reagents. Reagents are applied in a volume of 20 µl, incubated for 3-18 hours in a humid chamber at room temperature, and washed under running cold tap water. After the last reagent of the iodinated antibody preparation () is washed (), the washed and dried AEDs autoradiate, i.e. placed in contact with an x-ray film for 18 h. The x-ray film is then developed

The method is explained with examples.

Example 1. Diagnosis of hepatitis B in the detection of HBsAg in the sera of patients

Antibodies to anti-HBsAg (anti-HB) in an amount of 200 ng in a volume of 20 µl are applied to the probe points. Incubate in a humid chamber at room temperature for 3 hours. FBE is washed under running tap water and dried with filter paper.

Apply the test specimen - a human re-run, undiluted in a volume of 20 µl. Incubate at 18 h for 18 h. The probe is washed and dried.

Apply t-anti-hNBz 0.02 μK in a volume of 20 μl. They are incubated under conditions for 5 hours, the probe is washed, dried out, and placed in contact with the X-ray film for 18 hours. The X-ray film is developed and visualized.

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Estimated results. Mark the numbers of the dark points corresponding to the samples containing -HBsAg. Thus, the samples that darken the X-ray film contain HBsAg, and the samples that do not darken the X-ray film do not contain HBs.4g.

Example 2. Diagnosis of hepatitis A by detection of the virus in feces,

Antibodies against hepatitis A virus (anti-NAU) in a volume of 20 µl are applied to points on the probe. Incubated in a humid chamber at room temperature for 3 h, the probe was washed under running tap water and dried with filter paper.

Apply the test sample - fecal extract in a volume of 20 µl. Incubate in a humid chamber at room temperature for 18 hours. The probe is washed and dried.

Nanos t l-anti-NAU 0.02 μK in a volume of 20 μl. Incubated in a humid chamber for 5 hours. The probe was washed, dried, and placed in contact with an x-ray film for 18 hours. X-ray film exhibits and visually evaluates the results.

The samples, which provide darkening of the x-ray film, contain the hepatitis A virus, the samples that do not darken the x-ray film, do not contain the hepatitis A virus.

Example 3. Diagnosis of hepatitis A by the appearance of class M (igM) antiviral antibodies in the sera of patients.

Antibodies against human antibodies of class M (anti-M) in a volume of 20 µl are applied to points on the probe. Incubate in a humid chamber at room temperature for 3 hours. The probe is washed under running tap water and dried with filter paper.

Apply the test sample — human serum to be analyzed in a volume of 20 μl. Incubated in a humid chamber at room temperature for 18 hours. The probe is washed and dried.

The solution containing the hepatitis A virus is applied: the virus-containing extract of the patient’s feces in a volume of 20 µl is incubated in a moist chamber at a specific temperature of 5 hours. The probe is washed and dried.

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Nanos t I-antenna-NAU 0.02 μK in Samples containing antibodies with a volume of 20 μl. Incubated, washed with hepatitis A virus class M,

and dried. A probe is placed on a con-provide by darkening X-ray with an X-ray film on 18 tsp of the film. Samples that do not provide an x-ray film washed and video-5 darkening the x-ray film,

visually evaluate the results. Antiviral antibodies do not contain.

Claims (1)

METHOD FOR DETERMINING VIRAL HEPATITUS MARKERS by conducting radioimmunological analysis on a solid phase and detecting the radioactivity of the immune complex, characterized in that, in order to simplify the method while maintaining its high sensitivity, a plastic film is used as the solid phase, and the radioactivity of the immune complex is detected autoradiographically.