METHOD OF BLOOD ANALYSIS OF HIDV-INFECTED PATIENTS Russian patent published in 2024 - IPC C12Q1/68 

FIELD: biotechnology and virology.

SUBSTANCE: method for analyzing the blood of HIDV-infected patients to determine mutations in HIDV genes associated with changes in the functional properties of the virus is proposed. The method includes obtaining the patient's proviral DNA, conducting the first round of PCR, in which 4 PCR reactions are performed in separate tubes with pairs of primers SEQ ID:1 and SEQ ID NO:2, SEQ ID:3 and SEQ ID NO:4, SEQ ID:5 and SEQ ID NO:6, SEQ ID:7 and SEQ ID NO:8, respectively, while all 4 PCR reactions have the same amplification parameters. A proviral DNA sample obtained from a patient is used as an experimental sample, a buffer solution similar to the buffer solution in the experimental sample is used as a negative control, and as For positive control, DNA containing the corresponding gene of the non-structural HIDV-1 protein of sub-subtype A6 is used. Then a second round of PCR is performed, in which 4 PCR reactions with pairs of primers SEQ ID:9 and SEQ ID NO:10, SEQ ID:11 and SEQ ID NO:12, SEQ ID are placed in separate tubes13 and SEQ ID NO:14, SEQ ID:15 and SEQ ID NO:16, respectively, while experimental and control PCR mixtures obtained after the first round of PCR are used as experimental samples, while all 4 PCR reactions have uniform amplification parameters. Next, Sanger sequencing is performed with pairs of primers SEQ ID:9 and SEQ ID NO:10, SEQ ID:11 and SEQ ID NO:12, SEQ ID:13 and SEQ ID NO:14, SEQ ID:15 and SEQ ID NO:16. The nucleotide sequences of genes of nonstructural proteins are collected HIDV-1 is a sub-subtype of A6 based on data obtained at the Sanger sequencing stage. The obtained sequences are analyzed, during which mutations in the genes of non-structural proteins of HIDV-1 sub-subtype A6 are detected and the results are interpreted, during which mutations in the genes of non-structural proteins associated with changes in the functional properties of the virus are detected.

EFFECT: method of blood analysis of HIDV-infected patients makes it possible to identify mutations in the genes of non-structural proteins of HIDV-1 sub-subtype A6 circulating in the territory of the Russian Federation associated with changes in the functional properties of the virus, which can affect the course of HIDV infection, including the risk of developing comorbid diseases in the patient.

1 cl, 5 dwg, 3 tbl, 3 ex