应用多重巢式PCR法快速鉴定广西HIV-1主要亚型

摘要

目的 建立快速鉴定广西HIV-1主要流行亚型CRF01_AE、CRF07_BC、CRF08_BC、B和C的多重巢式PCR方法.方法 针对HIV-1 gag基因设计CRF01_AE、CRF07_BC、CRF08_BC、B和C亚型特异性引物,建立多重巢式PCR法,用该法鉴定来自广西的HIV-1毒株的亚型,并与基因测序法的鉴定结果比较.结果 多重巢式PCR法能正确鉴别5种已知亚型的样本,10份HIV阴性样本均无扩增,在72份未知亚型样本中,正确鉴定出66份,分别属于CRF01_AE、CRF07_BC、CRF08_BC、B亚型,多重巢式PCR法的灵敏度为91.7%,特异度达100%.结论 多重巢式PCR法能准确鉴定广西CRF01_AE、CRF07_BC、CRF08_BC和B亚型毒株,是一种简便、快速、低成本的亚型鉴定方法.%Objective To establish a rapid nested multiplex PCR assay for subtyping HIV-1 CRF01_AE,CRF07_BC,CRF08_BC,B,and C strains prevailing in Guangxi.Method Subtype-specific primers were designed for these subtypes based on their gag sequences.The subtypes of HIV-1 samples from Gmmgxi were determined by nested multiplex PCR and DNA sequencing and phylogenetic analysis,respectively,and then the sensitivity and the specificity of nested multiplex PCR were calcuhted.Results Nested multiplex PCR could correctly classify the 5 known-subtype samples,and were not reactive to all HIV-negative samples.Of the 72 HIV-positive samples,66 were correctly identified as CRFO1_AE,CRF07_BC,CRF08_BC,and B by this assay,giving a sensitivity of 91.7%(66/72),and a specificity of 100%.Conclusion This assay is a simple,fast,and cost-effective subtyping method for HIV-1 CRF01_AE,CRF07_BC,CRF08_BC,and B strains prevailing in Guangxi.