摘要目的 分析黑龙江省HIV-1感染者中流行株CRF01_AE env基因序列特征以及成簇特性.方法 应用巢式聚合酶链反应(Nested polymerase chain reaction,Nested-PCR)对env C2-C4区核苷酸序列进行扩增.用MEGA软件进行系统进化分析,然后对156份CRF01 _AE重组型HIV-1毒株env基因V3环及邻近区域氨基酸特征分析以及生物信息学分析.结果 156份样本中绝大多数样本与中国黑龙江、吉林、辽宁的代表毒株聚成一簇,还有一小部分样本与来自中国云南以及泰国代表株90CM240、90CM235聚成一簇.共发现6种V3环顶端四肽类型,主要类型为GPGQ、GPGR,此外还有GLGR、GQGR、GPGK、GPGS.根据V3环的氨基酸分析可预测HIV-1辅助受体的使用情况,其中121份(77.6％)使用趋化因子受体5(chemokine reseptor,CCR5)作为辅助受体,没有可被预测为使用趋化因子受体4[chemokine(c-x-c matif)-receptor 4,CXCR4]为辅助受体的毒株,35份(22.4％)样本不能被预测.以bootstrap≥90％、基因距离≤0.045作为界定传播簇的参数,共形成16条传播簇,其中1条传播簇有9条序列.结论 黑龙江省HIV-1流行株CRF01_AE V3环顶端四肽具有较大的变异性,大部分毒株呈现巨噬细胞嗜性.CRF01_AE毒株形成的传播簇规模较小,成簇序列数也仅达到四分之一.

abstractsObjective To analysis the sequence characheristics and clustering of env gene among HIV-1 prevalent strain CRF01_AE in Heilongjiang province.Method Nested polymerase chain reaction was used to amplify the env C2-G4 region nucleotide sequence.Phylogenetic analysis was carried out by MEGA.Then the amino acid characteristics and bioinformatics of the env V3 loop and adjacent regions of 156 CRF01_AE strains of recombinant HIV-1 were analyzed.Results Phylogenetic tree analysis showed that majority of the 156 samples clustered together with the representative strains from Heilongjiang,Jilin and Liaoning,a small number of samples clustered with representative strains from Yunnan and Thailand.The analysis of V3 loop amino sequence revealed that the CRF01 _AE strains had six types of V3 loop tetra peptide:GPGQ、GPGR、GLGR、GQGR、GPGK and GPGS.The main types were GPGQ and GPGR.Predictions for the potential use of coreceptors on the basis of the critical amino acids within V3 loop showed that 121 (77.6％) of the CRF01_AE strains utilized chemokine receptor 5 (CCR5) and none of the CRF01_AE strains utilized chemokine (c-x-c motif)receptor(CXCR4),while 35 (22.4％) could not be predicted.In the context of bootstrap ≥90％ and gene distance ≤0.045,16 transmission cluster were found and one of them contained 9 sequences.Conclusion As the dominate strains among HIV-1 in Heilongjiang province,CRF01_AE V3 loop has great variability and the majority of those currently circulating in Heilongjiang are likely to be non-syncytium inducing(NSI).The proportion of the transmission cluster is comperatively small and the cluster sequence number is only quartered.