CAJ | 학술논문

目的：探索应用单基因组扩增分析HIV-1准种基因变异。方法将深圳2010年6个基因测序出现重叠峰的样本RNA稀释至单拷贝，采用一步法RT-PCR套式扩增，扩增产物纯化后进行基因测序，应用Mega 4.02多功能基因分析软件分析不同准种序列的基因距离，构建进化树模型。同时，测定样本的CD4＋T淋巴细胞浓度和病毒载量。结果同一感染者体内HIV-1准种在进化树上形成了各自独立的簇，有些簇存在明显的亚簇，表明某些病毒株在复制中具有竞争优势。6个样本HIV准种的基因距离介于0.008与0.06之间，与感染持续时间和病毒载量有关。结论单基因组扩增法可以很好地分辨出HIV-1准种的基因变异，对于HIV-1准种基因距离的分析可以判断感染时间和分析免疫逃逸机制，其相关影响因素尚需更大样本的分析。
목적：탐색응용단기인조확증분석HIV-1준충기인변이。방법장심수2010년6개기인측서출현중첩봉적양본RNA희석지단고패，채용일보법RT-PCR투식확증，확증산물순화후진행기인측서，응용Mega 4.02다공능기인분석연건분석불동준충서렬적기인거리，구건진화수모형。동시，측정양본적CD4＋T림파세포농도화병독재량。결과동일감염자체내HIV-1준충재진화수상형성료각자독립적족，유사족존재명현적아족，표명모사병독주재복제중구유경쟁우세。6개양본HIV준충적기인거리개우0.008여0.06지간，여감염지속시간화병독재량유관。결론단기인조확증법가이흔호지분변출HIV-1준충적기인변이，대우HIV-1준충기인거리적분석가이판단감염시간화분석면역도일궤제，기상관영향인소상수경대양본적분석。
Objective To establish the single genome amplification ( SGA) method and analyze the quasispecies in HIV-infected patients.Methods All 6 sample RNA acquired in 2010 in Shenzhen and genetic sequenced as overlap peaks were extracted and diluted to a single copy , nest-PCR after one step RT-PCR was employed to amplify HIV-1 genome , and then PCR products was purified and sequenced.Mega 4.02 software was used to analyze the genetic distance among HIV-1 quasispecies , and phylogenetic tree was constructed.Results Our data showed that viral sequences derived from different patients were grouped into different clusters.Subcluster was also observed in several clusters , indicating these existed competition and preferential replication of certain viral strains.The genetic distance within one cluster of 6 samples were between 0.008 and 0.06, it was likely to associate with the duration since infection and viral load.Conclusion SGA is a useful approach to gain information on quasispecies , the genetic distance within one cluster may help to determine the infection time and immune escaping.The analysis of related affecting factors need more samples.