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METHOD TO EVALUATE THE SENSITIVITY OF HIV VARIANTS TO DRUGS ABLE TO INHIBIT THE HIV PROTEASE
METHOD TO EVALUATE THE SENSITIVITY OF HIV VARIANTS TO DRUGS ABLE TO INHIBIT THE HIV PROTEASE
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机译:评价HIV变种对能够抑制HIV蛋白酶药物的敏感性的方法
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摘要
A method to evaluate the sensitivity of an HIV variant from a biological sample to at least one drug able to inhibit the HIV protease is disclosed. The method comprises the steps of: a) amplifying the nucleic acid encoding the HIV protease from the biological sample, to obtain an amplified HIV protease coding sequence; b) cloning the amplified HIV protease coding sequence into a unique restriction site of a modified HIV molecular clone, wherein said modified HIV molecular clone is modified at least by deleting the protease coding sequence and by replacing it with the unique restriction site, to obtain a recombinant HIV molecular clone comprising and expressing the HIV protease coding sequence; c) introducing the recombinant HIV molecular clone comprising and expressing the HIV protease coding sequence into cultured cells, to obtain recombinant HIV transfected cells; d) measuring the level of HIV p24 antigen into the cell supernatant of the recombinant HIV transfected cells both in the presence and in the absence of the drug able to inhibit the HIV protease. The method is suitable for any HIV variant, both HIV-1 or HIV-2 variants. Diagnostic kits are also disclosed.
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