应用TaqMan荧光定量PCR技术建立HIV前病毒检测方法

Establishment of Real-time Fluorescence Polymerase Chain Reaction Based on the TaqMan Probes for Detection of HIV Provirus

目的建立HIV前病毒荧光定量PCR检测方法。方法常规培养8E5/LAV细胞,收集后计数,提取HIV前病毒核酸,运用实时荧光定量PCR法扩增晚期逆转录基因片段(HIVFL)。结果建立的实时荧光定量PCR技术检测灵敏度可达1拷贝,扩增结果稳定可靠。结论成功建立了HIV前病毒检测方法,为今后筛选和鉴定潜伏感染细胞以及评价抗-HIV药物的治疗效果奠定基础。

Objective To establish real-time fluorescence polymerase chain reaction（PCR） for HIV provirus detection. Methods HIV-1 provirus DNA was extracted after 8E5 cells were quantified. A fragment of late reverse transcripts （full-length） was tested by real-time fluorescence PCR. Results The quantitative real-time PCR assay was sensitive and specific. 1 copy of HIV FL could be consistently detected using this assay. Conclusion The method of HIV provirus detection was established successfully, which lay the foundation for further study on screening and verifying latent infected cells, evaluating the therapeutic efficacy of antiretroviral treatment.