CAJ | 학술논문

目的分析新疆地区HIV-1感染者中流行的单一亚型CRF07_BC毒株的gag基因遗传特性.方法对35例CRF07_BC亚型的HIV-1感染者分别于2011年9月(T1时间点)、2012年5月(T2时间点)、2013年2月(T3时间点)和2013年10月(T4时间点)4个时间点采集全血标本.为排除不同程度免疫功能对HIV-1基因变异的影响,使用流式细胞仪进行CD4+T淋巴细胞计数,并按照CD4+T淋巴细胞＜200/μL(重度免疫抑制)、200～500/μL(中度免疫抑制)和＞500/μL(无免疫抑制)分为不同免疫功能的3组.抽提标本RNA,套式PCR扩增gag基因片段,最终将3组研究对象得到的4次序列进行基因离散率和进化树分析,并使用BEAST软件估算该地区CRF07_BC流行毒株的最近共.同祖先株形成时间和平均进化速率.为排除HARRT对HIV-1基因变异影响,对HARRT组和非HARRT组进行基因离散率分析.结果 3组不同免疫功能的研究对象均呈现出在T1、T2、T3、T4时间点的组内和组间基因离散率依次升高的趋势.HARRT组和非HARRT组也均呈现出在T1、T2、T3、T4时间点的组内和组间基因离散率依次升高的趋势.进化树分析显示,同一样本的几次序列在进化树上聚集成一簇,大部分毒株均在T4时间点上独立分出一支;其最近共同祖先株形成时间为10.9年[95％最高后验密度(HPD):2.0～22.2年],平均进化速率为1.34×101替换·位点1·年-1(95％HPD:5.16×10-4～2.23×10-3替换·位点-1·年1).结论新疆地区HIV-1感染者CRF07_BC亚型毒株会随着在一个地区流行时间的延长,加大该地区同一亚型内的基因离散率;CRF07_BC单一亚型内病毒株的gag基因进化速率相对文献报道的略慢.
목적 분석신강지구HIV-1감염자중류행적단일아형CRF07_BC독주적gag기인유전특성.방법 대35례CRF07_BC아형적HIV-1감염자분별우2011년9월(T1시간점)、2012년5월(T2시간점)、2013년2월(T3시간점)화2013년10월(T4시간점)4개시간점채집전혈표본.위배제불동정도면역공능대HIV-1기인변이적영향,사용류식세포의진행CD4+T림파세포계수,병안조CD4+T림파세포＜200/μL(중도면역억제)、200～500/μL(중도면역억제)화＞500/μL(무면역억제)분위불동면역공능적3조.추제표본RNA,투식PCR확증gag기인편단,최종장3조연구대상득도적4차서렬진행기인리산솔화진화수분석,병사용BEAST연건고산해지구CRF07_BC류행독주적최근공.동조선주형성시간화평균진화속솔.위배제HARRT대HIV-1기인변이영향,대HARRT조화비HARRT조진행기인리산솔분석.결과 3조불동면역공능적연구대상균정현출재T1、T2、T3、T4시간점적조내화조간기인리산솔의차승고적추세.HARRT조화비HARRT조야균정현출재T1、T2、T3、T4시간점적조내화조간기인리산솔의차승고적추세.진화수분석현시,동일양본적궤차서렬재진화수상취집성일족,대부분독주균재T4시간점상독립분출일지;기최근공동조선주형성시간위10.9년[95％최고후험밀도(HPD):2.0～22.2년],평균진화속솔위1.34×101체환·위점1·년-1(95％HPD:5.16×10-4～2.23×10-3체환·위점-1·년1).결론 신강지구HIV-1감염자CRF07_BC아형독주회수착재일개지구류행시간적연장,가대해지구동일아형내적기인리산솔;CRF07_BC단일아형내병독주적gag기인진화속솔상대문헌보도적략만.
Objective To analyze the genetic dynamics of prevailing single subtype CRF07_BC in human immunodeficiency virus-1 (HIV-1) infected patients in Xinjiang.Methods Blood samples were collected at four time points,including in 2011 September (T1 time point),in 2012 May (T2 time point),in 2013 February (T3 time point) and in 2013 October (T4 time point),from 35 HIV-1 infected patients of subtype CRF07_BC in Xinjiang.CD4+ T lymphocyte count was conducted by using flow cytometry to eliminate the effect of immune function on HIV-1 gene mutation.All cases were divided into three groups according to CD4+ T lymphocyte counts＜200/μL (severe immune suppression),200-500/μL (moderate immune suppression) and ＞ 500/μL (nonimmuno suppression).Then HIV-1 RNA was extracted and HIV-1 gag gene was amplified by nested polymerase chain reaction (nPCR).The gag gene was sequenced by gene distance and phylogenetic analyses.The time to the formulation of the most recent common ancestor (tMRCA) and evolutionary rate of subtype CRF07_BC in Xinjiang were estimated by software Bayesian evolutionary analysis by sampling trees (BEAST).Analysis of gene distance was conducted in highly active antiretroviral therapy (HAART) group and nonHAART group to eliminate the effect of HAART on the HIV-1 mutation.Results The gene distance within 3 groups at T1 time point,T2 time point,T3 time point and T4 time point showed an increasing trend respectively.Gene distance in HAART group and nonHAART group also showed the similar trend.Phylogenetic tree showed a cluster of sequences of the same samples and the branches were split into a new one at T4 time point.tMRCA was 10.9 years (95 ％ highest posterior density [HPD]:2.0-22.2 years).Evolutionary rate was estimated to be 1.34 × 10-3 substitutions · site-1 · year-1 (95 ％ HPD:5.16 × 10-4-2.23 × 10-3 substitutions · site 1 · year-1) substitutions per site per year.Conclusions With the viruses prevailing in Xinjiang,the gene distance increases in the single subtype CRF07_BC in different immune function groups.Evolutionary rate of gag gene of subtype CRF07_BC is slower than that of reference.