HIV-1 resistance strain and use thereof
A technology for drug resistance and virus strains, applied in the direction of viruses/phages, medical materials derived from viruses/phages, microorganisms, etc., can solve the problems of lack of systematic research and constraints on HIV drug resistance
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Embodiment 1、3
[0008] Embodiment 1, the cultivation and screening of 3TC HIV-1 drug-resistant strains
[0009] 1. Experimental materials
[0010] 1. Virus: HIV-1 CNHN24 strain, which was isolated from peripheral blood lymphocytes of AIDS patients in Henan, China in 2002, Genbank registration number AY180905, genotype is B' subtype, and the band type detected by HIV antibody immunoblotting is gp160, gp120, p66, p55, gp41, p24, p17.
[0011] 2. Cells: MT4 cells, cultured and subcultured with RPMI1640 medium containing 10% calf serum.
[0012] 3. Screening drugs: 3TC drugs are Heprudine Lamivudine Tablets produced by GlaxoSmithKline Pharmaceuticals, each containing 100 mg of Lamivudine. Dissolve 100mg of 3TC in 50ml of RPMI1640 culture medium, and centrifugally filter to obtain 8.7mM 3TC solution, which should be diluted to the desired concentration before use.
[0013] 4. Cell culture medium: RPMI1640 cell culture medium containing 10% calf serum.
[0014] 2. Cultivation of 3TC HIV-1 drug-...
Embodiment 2
[0032] Embodiment 2, application of 3TC HIV-1 drug-resistant strains to screen anti-HIV drugs
[0033] 1. Experimental materials
[0034] 1. The test drug DCP 961 ( *J. Med. Chem. 2000, 43, 2019-2030. Inhibition of Clinically Relevant Mutant Variants of HIV-1 by Quinazolinone Non-Nucleoside Reverse Transcriptase Inhibitors. Jeffrey W. Corbett, Soo S. Ko, James D. Rodgers, et al)
[0035] Physical and chemical properties: white powder
[0036] 2. Positive control drug AZT
[0037] Provider: SIGMA, batch number: 021033
[0038] 3. Cells
[0039] MT-4, preserved by the Army AIDS Testing Center of the Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, was cultured and subcultured with RPMI 1640 medium containing 10% fetal bovine serum.
[0040] 4. Viruses
Embodiment 1
[0041] The 3TC HIV-1 drug-resistant strain obtained in Example 1.
[0042] 5. Basic conditions of the experiment
[0043] The anti-HIV-1 activity experiment was carried out in a biosafety level 3 laboratory.
[0044] 2. Drug screening process
[0045] 1. The infectivity titer (TCID) of virus to MT-4 cells was determined by CPE method 50 )
[0046] MT-4 cells were seeded in a 96-well plate, 100 μl per well, containing 2.5×10 cells 4 cells / ml, the virus stock solution was diluted 10 times with the culture medium, and a total of 9 concentrations were added to the cell wells, 4 wells per concentration, 37 ° C, 5% CO 2 Incubator cultivation. Cytopathic changes (CPE) were observed daily with an inverted microscope. Cytopathic changes below 25% are +, 26-50% lesions are ++, 51-75% lesions are +++, and 76-100% lesions are ++++. Cytopathic extent (CPE) was recorded. The infectious titer (TCID) of the virus was calculated by the Reed-Muench method 50 ).
[0047] 2. MTT method ...
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