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A HIV-1crf01_ae subtype strain and its application

A technology of HIV-1GX002 and HIV-1, applied in the field of HIV-1CRF01_AE subtype strains, can solve problems such as constraints and lack of high replication ability

Active Publication Date: 2018-08-14
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no standard HIV-1 clinical isolate with high replication ability used in the evaluation of new AIDS drugs. On the other hand, the research on HIV-1 replication ability in my country is also limited by the lack of standard HIV-1 clinical isolates with high replication ability restricted

Method used

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  • A HIV-1crf01_ae subtype strain and its application
  • A HIV-1crf01_ae subtype strain and its application
  • A HIV-1crf01_ae subtype strain and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1. Isolation, cultivation and determination of full-length genome sequence of GX002

[0053] 1. Isolation and subculture of HIV-1 CRF01_AE subtype strain GX002 with high replication ability

[0054] (1) Centrifuge the whole blood obtained from an AIDS patient in Guangxi Zhuang Autonomous Region, China, at 250g for 8 minutes to separate the plasma from the blood cells, discard the upper layer of plasma, suck the lower layer of blood cells into a 50ml centrifuge tube, and add a physiological volume equal to that of the plasma Dilute and mix with saline to obtain a diluted blood cell suspension;

[0055] (2) Slowly add the separation solution HISTOPAQUE-1077 and the diluted blood cells to another new centrifuge tube at a volume ratio of 1:1. A clear separation liquid surface is formed between the blood cell suspension and the separation liquid;

[0056] (3) After the centrifuge tube in step (2) was centrifuged at 400g for 30 minutes, the liquid surface was divide...

Embodiment 2

[0084] Example 2, Application of GX002 strain to evaluate the efficacy of anti-HIV-1 drugs

[0085] 1. The cytopathic effect (CPE) observation method was used to determine the infectivity titer of the virus to cells (TCID 50 )

[0086] MT-4 cells were seeded in a 96-well plate, 100 μl per well, and the cell concentration was 5×10 4 / ml, the GX002 virus stock solution was diluted 10-fold successively with culture medium (containing 10% fetal bovine serum, 100 IU / ml penicillin, 100 μg / ml streptomycin RPMI1640 medium) and added 8 concentration gradients successively. Into the cell wells, 4 parallel wells per concentration, 37°C, 5% CO 2 Incubator cultivation. Cytopathic effect (CPE) was observed with an inverted microscope every day. The infectious titer (TCID) of GX002 strain was calculated by Reed-Muench method 50 ).

[0087] 2. MTT method was used to determine the half toxic concentration (TC) of the tested drug on MT-4 cells, H9 cells and human peripheral blood mononucl...

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Abstract

The invention discloses a HIV-1 CRF01-AE subtype (Human immunodeficiency virus type I subtype CRF01-AE) virus strain and application thereof, the (Human immunodeficiency virus type I subtype CRF01-AE) virus has the preservation number of CGMCC No.8787. The clinical isolate GX002 of the HIV-1 CRF01-AE subtype can be widely used in screening of drugs for treatment of AIDS (acquired immune deficiency syndrome) and HIV-1 replication capacity related researches, and has important application value and significance on the study of the prevention and treatment of AIDS.

Description

technical field [0001] The invention relates to an HIV-1CRF01_AE subtype strain and application thereof, belonging to the field of biotechnology. Background technique [0002] Since the Acquired Immunodeficiency Syndrome (AIDS) caused by Human Immunodeficiency Virus (Human Immunodeficiency Virus, HIV) was introduced into China in 1985, the scope of its spread in China has gradually increased, and the rate of transmission has been increasing. quick. Its popularity has gone through the introduction period and the diffusion period, and is currently entering a period of rapid growth. According to the epidemic report of the China Center for STD and AIDS Prevention and Control in the first quarter of 2014, as of March 31, 2014, there were 455,352 existing or HIV / AIDS cases nationwide, 140,750 deaths, 275,631 living HIV infections, and 179,721 AIDS patients. . Sexual transmission becomes the main route of transmission, accounting for about 90% of all infections (both heterosexua...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12Q1/70C12Q1/18C12R1/93
Inventor 韩婧婉刘思扬庄道民李韩平梁淑家李敬云
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI