HIV-1逆转录过程及应用PCR检测

Process of HIV-1 reverse transcription and its detection by using PCR

人类免疫缺陷病毒(human immunodeficiency virus,HIV)作为逆转录病毒,在进入宿主细胞以后其正链RNA基因组就在逆转录酶的作用下开始逆转录合成双链的DNA。HIV-1逆转录的过程大致包括以下几个步骤:负链强力终止DNA的起始合成,负链强力终止DNA的链转移,正链DNA的合成起始,正链强力终止DNA的链转移和最终完整双链DNA的合成。逆转录是HIV-1复制的重要环节,现已成为抗HIV-1药物研究开发的重要靶标。聚合酶链反应(polymerase chain reaction,PCR)可用来检测HIV-1的逆转录过程,设计引物扩增HIV-1基因组R/U5序列、U3序列、U5/PBS序列和LTR与Gag之间的序列,根据PCR结果可相应判断出HIV-1的逆转录是否起始、是否进行了负链的链转移、正链的起始和双链DNA的完整合成。本文将主要介绍逆转录过程以及应用PCR的检测方法。

Human immunodeficiency virus （HIV） is a retrovirus, belongs to Lentiviridae family. As long as viral genetic material entering into host cytoplasm, double-strand DNAs synthesis occurs which is catalyzed by reverse transcriptase （RT） with viral plus-strand RNA as template. This reverse transcription is a key link of HIV-1 life cycle and an important target for anti-HIV drug development. The process of reverse transcription can be divided into several steps： formation of minus-strand strong-stop DNA; the first translocation; initiation of plus-strand DNA synthesis; and, the second translocation and the completion of both strands. These steps can be detected individually by using polymerase chain reaction （PCR） according to the amplified products on the region of R/U5, U3, U5/PBS and the sequence between LTR and Gag. In this review, we summarize the principle for detecting stages of HIV-1 reverse transcription by using PCR.