首页> 外国专利> 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

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

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.
机译:公开了一种评估来自生物样品的HIV变体对至少一种能够抑制HIV蛋白酶的药物的敏感性的方法。该方法包括以下步骤:a)从生物样品中扩增编码HIV蛋白酶的核酸,以获得扩增的HIV蛋白酶编码序列; b)将扩增的HIV蛋白酶编码序列克隆到修饰的HIV分子克隆的独特限制性位点,其中所述修饰的HIV分子克隆至少通过缺失蛋白酶编码序列并通过用独特的限制性位点替换而被修饰,以获得重组HIV分子克隆,其包含并表达HIV蛋白酶编码序列; c)将包含并表达HIV蛋白酶编码序列的重组HIV分子克隆导入培养细胞中,以获得重组HIV转染的细胞; d)在存在和不存在能够抑制HIV蛋白酶的药物的情况下,测量进入重组HIV转染细胞的细胞上清液中的HIV p24抗原的水平。该方法适用于任何HIV变体,HIV-1或HIV-2变体。诊断试剂盒也被公开。

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