Emery S, Bodrug S, Richardson B A, Giachetti C, Bott M A, Panteleeff D, Jagodzinski L L, Michael N L, Nduati R, Bwayo J, Kreiss J K, Overbaugh J
Division of Human Biology, Fred Hutchinson Cancer Research Center, University of Washington, Seattle, Washington 98109, USA.
J Clin Microbiol. 2000 Jul;38(7):2688-95. doi: 10.1128/JCM.38.7.2688-2695.2000.
Accurate and sensitive quantification of human immunodeficiency virus type 1 (HIV-1) RNA has been invaluable as a marker for disease prognosis and for clinical monitoring of HIV-1 disease. The first generation of commercially available HIV-1 RNA tests were optimized to detect the predominant HIV-1 subtype found in North America and Europe, subtype B. However, these tests are frequently suboptimal in detecting HIV-1 genetic forms or subtypes found in other parts of the world. The goal of the present study was to evaluate the performance of a new viral load assay with non-subtype B viruses. A transcription-mediated amplification method for detection and quantitation of diverse HIV-1 subtypes, called the Gen-Probe HIV-1 viral load assay, is under development. In this study we examined the performance of the Gen-Probe HIV-1 viral load assay relative to that of the commonly used commercial HIV-1 RNA assays using a panel of primary isolates from Kenya. For comparison, we included several subtype B cloned viruses, and we quantified each virus using an in-house quantitative-competitive reverse transcriptase PCR (QC-RT-PCR) method and gag(p24) antigen capture. The Gen-Probe HIV-1 viral load assay and a version of the Roche AMPLICOR HIV-1 MONITOR test (version 1.5) that was designed to detect a broader range of subtypes were both sensitive for the quantification of Kenyan primary isolates, which represented subtype A, C, and D viruses. The Gen-Probe HIV-1 viral load assay was more sensitive for the majority of viruses than the Roche AMPLICOR HIV-1 MONITOR test version 1.0, the Bayer Quantiplex HIV RNA 3.0 assay, or a QC-RT-PCR method in use in our laboratory, suggesting that it provides a useful method for quantifying HIV-1 RNAs from diverse parts of the world, including Africa.
准确且灵敏地定量检测1型人类免疫缺陷病毒（HIV-1）RNA，作为疾病预后的标志物以及HIV-1疾病临床监测的手段，具有重要价值。第一代市售的HIV-1 RNA检测方法经过优化，用于检测在北美和欧洲发现的主要HIV-1亚型，即B亚型。然而，这些检测方法在检测世界其他地区发现的HIV-1基因形式或亚型时，常常不尽人意。本研究的目的是评估一种新型病毒载量检测方法对非B亚型病毒的检测性能。一种用于检测和定量多种HIV-1亚型的转录介导扩增方法，即Gen-Probe HIV-1病毒载量检测方法，正在研发中。在本研究中，我们使用一组来自肯尼亚的原始病毒分离株，检测了Gen-Probe HIV-1病毒载量检测方法相对于常用的市售HIV-1 RNA检测方法的性能。为作比较，我们纳入了几种B亚型克隆病毒，并使用内部定量竞争逆转录酶PCR（QC-RT-PCR）方法和gag（p24）抗原捕获法对每种病毒进行定量。Gen-Probe HIV-1病毒载量检测方法以及罗氏AMPLICOR HIV-1 MONITOR检测方法（1.5版）的一个旨在检测更广泛亚型范围的版本，对代表A、C和D亚型病毒的肯尼亚原始病毒分离株的定量检测均很灵敏。与罗氏AMPLICOR HIV-1 MONITOR检测方法1.0版、拜耳Quantiplex HIV RNA 3.0检测方法或我们实验室使用的一种QC-RT-PCR方法相比，Gen-Probe HIV-1病毒载量检测方法对大多数病毒更为灵敏，这表明它为定量来自包括非洲在内的世界不同地区的HIV-1 RNA提供了一种有用的方法。
