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目的对在深圳发现的一例gag和env基因片段亚型不一致的HIV感染者样本进行近全长基因组扩增和序列分析,以了解其重组特征并分析可能的来源。方法利用近末端稀释法分两段扩增病毒近全长基因组并测定序列,使用Sim Plot 3.5软件分析全长基因组序列中的重组断点,并采用分片段构建进化树的方法确认重组断点。对长度超过300 bp的片段与相应亚型的全部近全长基因组序列构建进化树,分析亲本毒株可能的来源。结果经扩增测序获得1条长度为8 975 bp的HIV-1近全长基因组序列。断点分析结果表明该序列由CRF01_AE和CRF07_BC重组而成,重组断点相应于HXB2的位置分别为1 185、1 817、8 782、9 043和9 216,其中片段Ⅰ、Ⅲ、Ⅴ来源于CRF01_AE毒株,片段Ⅱ、Ⅳ、Ⅵ来源于CRF07_BC毒株。亲本毒株来源分析表明,与片段Ⅱ成簇的CRF07_BC序列主要来自北方男男同性传播人群,Bootstrap值为74%,片段Ⅲ与CRF01_AE的簇5病毒成簇,其中多数序列来自北方男男同性传播人群,Bootstrap值为100%。结论本文获得了一株CRF01_AE和CRF07_BC重组形成的独特型重组病毒,其亲本病毒与我国北方男男同性恋人群流行的毒株同源,有可能来源于深圳本地性传播人群中流行的病毒。
OBJECTIVE: To perform near full-length genomic amplification and sequence analysis on a sample of HIV-infected individuals found in Shenzhen that are not consistent with subtypes of gag and env gene fragments to understand their recombination characteristics and to analyze possible sources. Methods Near-end dilution was used to amplify the full-length genome of the virus in two stages and the sequence was determined. Simian plot 3.5 software was used to analyze the recombination breakpoints in the full-length genome sequence and the fragment was constructed by using the segmented construction phylogenetic tree. Phylogenetic trees were constructed for all nearly full-length genomic sequences of fragments longer than 300 bp and corresponding subtypes, and the possible sources of the parental strains were analyzed. Results A full-length HIV-1 genome sequence of 8 975 bp was obtained by sequencing. The breakpoint analysis showed that the sequence was recombined by CRF01_AE and CRF07_BC. The positions of the recombination breakpoint corresponding to HXB2 were 1 185, 1 817, 8 782, 9 043 and 9 216, respectively. The fragments Ⅰ, Ⅲ and Ⅴ were derived from CRF01_AE Strain, fragment Ⅱ, Ⅳ, Ⅵ from CRF07_BC strains. Analysis of the origin of the virulent strains showed that the sequence of CRF07_BC clustered with fragment Ⅱ mainly came from the same-sex gay population in northern China. The Bootstrap value was 74%, and the cluster 5 virus of fragment Ⅲ and CRF01_AE were clustered. Most of the sequences were from homosexual transmission The crowd, Bootstrap value is 100%. Conclusions This study obtained a unique recombinant recombinant virus of CRF01_AE and CRF07_BC. Its parental virus is homologous with the strains of the gay male homosexuals in northern China, possibly from the viruses circulating in the local sexually transmitted population in Shenzhen.