Real-time PCR assay of individual human immunodeficiency virus type 1 variants in coinfected human lymphoid tissues.
Real-time PCR assay of individual human immunodeficiency virus type 1 variants in coinfected human lymphoid tissues.
复制标题
对共感染的人淋巴组织中的个体人类免疫缺陷病毒 1 型变种进行实时 PCR 测定。
DOI：
10.1128/jcm.41.5.2126-2131.2003
复制
发表时间：
2003
影响因子：
9.4
通讯作者：
Margolis,Leonid
中科院分区：
文献类型：
--
作者：
Ito,Yoshinori;Grivel,Jean-Charles;Margolis,Leonid
在感染过程中，人类免疫缺陷病毒1型（HIV-1）发生突变，分化成一个“群”的病毒准种，和CCR 5或CXCR 4利用准种的优势是密切相关的疾病进展的模式。病毒群中利用CCR 5和CXCR 4的病毒的定量在研究这种现象的机制中是重要的。在这里，我们报告了一个新的实时PCR为基础的方法学的复制个别CCR 5和CXCR 4利用变异的评估。该测定具有高度的可重复性，变异系数<3%，并且即使当它们在混合物中的差异为2个数量级时，它也能准确地估计病毒特异性RNA拷贝的数量。我们证明，复制CCR 5和CXCR 4利用的变体可以在实验共感染的人淋巴组织中进行评估和区分。我们开发的检测方法可能有助于研究感染HIV-1的人体组织中病毒群中R5到X4转换的机制。
In the course of infection, human immunodeficiency virus type 1 (HIV-1) mutates, diverging into a “swarm” of viral quasispecies, and the predominance of CCR5- or CXCR4-utilizing quasispecies is strongly associated with the pattern of disease progression. Quantification of CCR5- and CXCR4-utilizing viruses in viral swarms is important in the investigation of the mechanisms of this phenomenon. Here, we report on a new real-time PCR-based methology for the evaluation of replication of individual CCR5- and CXCR4-utilizing variants. The assay is highly reproducible, with a coefficient of variation of <3%, and it accurately estimates the numbers of virus-specific RNA copies even when their difference in the mixture is 2 orders of magnitude. We demonstrate that replications of CCR5- and CXCR4-utilizing variants can be evaluated and distinguished in experimentally coinfected human lymphoid tissue. The assay we developed may facilitate study of the mechanisms of the R5-to-X4 switch in viral swarms in human tissues infected with HIV-1.