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目的研究云南省目前主要流行的艾滋病病毒1型(HIV-1)的独特型重组(URFs)毒株的分子变异特点。方法收集2012-2014年云南省14个地市,800例在定点医院随访的HIV感染者或艾滋病患者的血样及流行病学调查信息,采用反转录聚合酶链反应(RT-PCR)的方法,分别扩增HIV-1的3′半分子和5′半分子。测序后,序列应用contig软件校对拼接,获得约9.0kb的近全长基因组,后用Genotyping、MEGA 6.06和BLAST工具软件确定毒株亚型,挑出新型独特型重组毒株,分析云南省HIV-1独特型重组毒株的基因变异及重组进化情况。结果共获得800例样本的103条近全长基因组,分析后挑出独特型重组毒株20个,分别来自于昆明、德宏、红河、昭通、大理和西双版纳地区。20个毒株之间的平均基因距离为(0.204±0.015),分别与来自泰国、印度、越南和日本的国际参考株的平均基因距离较为接近。云南省HIV-1独特型重组株呈现3种基因重组模式,主要为以C亚型为骨架的11株,其次是CRF0l_AE为骨架的7株,和B(B′)为骨架的2株。从重组区域上看,env、nef和rev基因是最易发生重组的基因区域。结论在2012-2014年间,云南省的HIV-1毒株呈现高度重组进化趋势,出现了大量的独特型重组毒株,且这些毒株与一些东南亚国家的毒株表现出高度同源性,其重组的类型和模式比较复杂,并且在全省范围内广泛传播开来,应密切监测流行趋势变化。
Objective To study the molecular variation of HIV-1 unique recombinant strains (URFs) in Yunnan Province. Methods Blood samples and epidemiological investigation were collected from HIV-infected or AIDS-infected patients in 14 cities and prefectures in Yunnan Province during the period of 2012-2014. 800 patients were followed-up in designated hospitals. Reverse transcription-polymerase chain reaction (RT-PCR) , Amplifying 3 ’and 5’ molecules of HIV-1, respectively. After sequencing, the sequences were contig-proofed by contig software to obtain the nearly full-length genome of about 9.0 kb. Genotyping, MEGA 6.06 and BLAST tools were used to identify the subtypes of the strains and a new unique type of recombinant strains were picked out to analyze HIV- 1 unique recombinant strains of the genetic variation and recombinant evolution. Results A total of 103 full-length genomes of 800 samples were obtained. Twenty unique recombinant strains were isolated from Kunming, Dehong, Honghe, Zhaotong, Dali and Xishuangbanna respectively. The average genetic distance between the 20 strains was (0.204 ± 0.015), which was close to that of the international reference strains from Thailand, India, Vietnam and Japan, respectively. There are three kinds of recombination patterns of HIV-1 unique recombinant strains in Yunnan Province, including 11 strains with C subtype as skeleton, 7 strains with CRF0l_AE as skeleton and 2 strains with B (B ’) as skeleton. From the perspective of recombination region, env, nef and rev genes are the most prone to recombination gene regions. Conclusions During 2012-2014, HIV-1 strains in Yunnan Province showed a highly evolutionary trend of evolution. A large number of unique recombinant strains appeared. These strains showed high homology with strains of some Southeast Asian countries. The types and patterns of reorganization are complex and spread widely across the province and should closely monitor changes in trends.