High-throughput human immunodeficiency virus type 1(HIV-1) full replication assay that includes HIV-1 vif as an antiviral target
High-throughput human immunodeficiency virus type 1(HIV-1) full replication assay that includes HIV-1 vif as an antiviral target
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DOI：
10.1128/aac.49.9.3833-3841.2005
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发表时间：
2005-09-01
影响因子：
4.9
通讯作者：
Blair, WS
中科院分区：
文献类型：
--
作者：
Cao, J;Isaacson, J;Blair, WS
抗病毒筛选已被证明可用于鉴定新型人类免疫缺陷病毒1型（HIV-1）抑制剂。在这项研究中，我们描述了一种HIV-1完全复制（HIV-1 Rep）检测方法，该方法结合了T细胞系中复制所需的所有靶标，包括HIV-1 Vif基因。HIV-1 Rep检测试剂盒旨在对晚期和早期抑制剂表现出最佳灵敏度，以最大限度地提高筛选中识别新型靶向抗病毒化合物的可能性。此外，HIV-1 Rep检测试剂盒的灵活性允许在不同T细胞系中快速评价抗病毒化合物对不同病毒株的抗病毒作用，而无需显著修改检测试剂盒形式。我们证明了HIV-1 Rep检测具有以下特征（例如，有利的Z ′值）与384孔格式中的高通量筛选相容。HIV-1 Rep测定的实用性在> 10（6）种化合物的高通量筛选中得到证实。据我们所知，这项研究代表了HIV-1抗病毒筛选的第一个例子，其中包括Vif作为功能靶标，并在工业规模上进行。
Antiviral screens have proved useful for the identification of novel human immunodeficiency virus type 1 (HIV-1) inhibitors. In this study, we describe an HIV-1 full replication (HIV-1 Rep) assay that incorporates all of the targets required for replication in T-cell lines, including the HIV-1 Vif gene. The HIV-1 Rep assay was designed to exhibit optimal sensitivity to late-stage as well as early-stage inhibitors to maximize the likelihood of identification of novel target antiviral compounds in a screen. In addition, the flexibility of the HIV-1 Rep assay allows the rapid evaluation of antiviral compounds against different virus strains in different T-cell lines without significant modification of the assay format. We demonstrate that the HIV-1 Rep assay exhibits characteristics (e.g., a favorable Z' value) compatible with high-throughput screening in a 384-well format. The utility of the HIV-1 Rep assay was demonstrated in a high-throughput screen of > 10(6) compounds. To our knowledge, this study represents the first example of an HIV-1 antiviral screen that includes Vif as a functional target and was executed on an industrial scale.