SU1768637A1 - Strain of human immunodeficient virus 1-type for preparation of diagnostic preparation - Google Patents

Description

The invention relates to the field of virology and biotechnology.

Known strains HIV-I / IIIB (USA), HIV-1 / CBL-4/85 (Great Britain), deposited in the State collection of viruses of the USSR. Due to their commercial use in countries where the virus is isolated, they are not available for use. Such domestic virus strains are not described in the literature (1,2).

The problem of AIDS diagnostics is extremely urgent for the Soviet health care. To conduct a mass survey of the population requires a significant number of diagnostic test systems of various types. The main component of most of the test systems used is the viral antigen. That is why the sensitivity and specificity of the produced diagnostics largely depend on the properties of the virus strain. In this regard, the use of viral strains endemic to the territory of the USSR in domestic test systems is of particular importance.

A new original strain of the human immunodeficiency virus HIV-1 / USSR, IV / YV-17/89, isolated on the territory of the USSR and intended for the preparation of diagnostic preparations for detecting HIV infection among the population, is proposed.

The virus strain is stored in the State Collection of Viruses of the Institute of Virology. D.I. Ivanovsky of the USSR Academy of Medical Sciences.

The aim of the present invention is to provide an original strain of human immunodeficiency virus with high infectious and antigenic activity, allowing it to be used for the preparation of diagnostic preparations for the detection of HIV-infected individuals among the population.

Identification of the strain HIV-1 / USSR, IV / YV-17/89 was carried out by a number of independent methods.

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1768637 A1

The virus strain was isolated from peripheral blood lymphocytes of an AIDS patient. When obtaining the strain, methods were used for co-cultivation of patient's lymphocytes with blood lymphocytes from a healthy donor, 5 stimulated by mitogen, as well as with human transplantable cell lines.

Reproduction. Strain; the virus replicates in peripheral blood lymphocytes, lymphoblastoid cell cultures 10 MT-4, CEM, Jurkat tat 3, V-937. Reproduction of the virus was accompanied by a characteristic cytopathic effect and syncytium formation. The infectious titer of the virus was determined by 50% tissue cytopathic dose of the virus on the model of the above cell cultures (Table 1),

Morphology. The method of electron microscopy revealed virions with a size of 100-120 nm with a morphology typical of human immuno-20 deficiency viruses of type 1.

Analysis of antigens.

Immunofluorescence. The study by the method of indirect immunofluorescence using the serum of the patient with AIDS 25, containing antibodies to HIV-1, showed the presence of HIV-1 antigens in 50-60% of infected cells.

Electrophoresis, immunoblot. The electrophoretic profile of the proteins of the strain corresponds to the protein spectrum of HIV-1.

The study in immunoblot allowed the conclusion that the virus has antigenic determinants characteristic of HIV-1: 120-160, 53/55. 41, 31.24.17 kD. 35

The identification of the strain was also carried out by means of point hybridization and determination of reverse transcriptase activity.

PRI me R 1. Diagnosis of antibodies in 40 HIV-1 by enzyme immunoassay

Material for research: Sera of HIV-1-infected persons, AIDS patients and fractions of immunoglobulins from sera of HIV-infected persons. Three strains were used as antigen. 45

HIV-1: reference strains IIIB, CB1_-4 and strain HIV-1 / IV / 17. The enzyme immunoassay was carried out according to the standard procedure (1). ,

The results of the enzyme immunoassay are presented in table. 2.

The results obtained show that the antigen preparation prepared from the proposed strain of HIV-1 / USSR, IV / IV-17, has the necessary antigenic determinants to detect virus-specific antibodies from the serum of HIV-1-infected individuals.

Comparative analysis of this strain with two reference ones in the enzyme-linked immunosorbent assay indicates its similar activity.

The proposed original strain of the HIV-1 / USSR virus, IV / IV-17 and diagnostic preparations prepared on its basis should find application in virological practice for the detection of antibodies in the sera of HIV-infected persons.

PRI me R 2. Diagnosis of antibodies to HIV-1 by immunoblot method.

Material for the study of antibodies to HIV-G is represented by 4 sera containing these antibodies and previously characterized in commercial test systems from DuPont and Biorad. The antigen used was the HIV-1 / IV-17 strain and the reference HIV1 / IIIB strain, purified according to the standard method. Immunoblot bands were also prepared according to the standard procedure (2).

The research results are presented in the table.

As a result of the studies, it was found that the antigen of the HIV1 / IV-17 strain allows the detection of antibodies to all HIV-1 polypeptides and can therefore be used to study the seroconversion of persons infected with HIV-1,

Claims (1)

Claim Human immunodeficiency virus strain 1-type GKV No. 4022 for the preparation of diagnostic preparations. Table! Investigation of the infectious activity of HIV strains using the Jurkat tat 3 cell model Caption, lg TCD 50 Strains of HIV-1 111B CBL-4 IV-17 6.5 ... 4.5 6.0 Table 2 Analysis of HIV-1 antigens by enzyme immunoassay Antibody preparations Virus antigens * HIV-1 / 111B HIV-1 / CBL-4 HIV-1 / IV-17 Serum of HIV-1 infected 16000 • 8000 4000 Fraction of immunoglobulins from serum of HIV-1-infected persons 32000 45 4000 8000 * - The reciprocal of the antibody titer Table 3 Study of antibodies to HIV-1 by immunoblot method Viral antigen Revealed antibodies HIV-1 / 111B 160 120 65 55 53 41 31 24 17 HIV-1 / IV-17 160 120 65 55 53 41 31 24 17