CAJ | 학술논문

目的探讨中国丙型肝炎患者中HCV的基因型分布类型,并进行HCV在中国流行病学历史分析,进而研究HCV在中国的分子流行病学和进化动力学特征.方法从423例丙型肝炎患者的血清中提取HCV RNA并进行cDNA反转录,扩增E1和NS5B两个基因区的核苷酸序列,排除不合格及不适合分析的病毒株序列,再对合格序列进行系统发育分析(phylogenetic analysis).运用BEAST软件中的Bayesian MCMC (Markov Chain Monte Carlo)算法来进行Coalescence分析,通过重构BSPs(Bayesian skyline plots)图回溯HCV的流行病学历史.结果 HCV分离株的基因分型如下:共包括6个基因型,12个基因亚型(1b:65.9％,6a:17.1％,2a:7.4％,3a:3.6％,3b:3.3％,6e:0.76％,1a,1c,2b,2f,4d以及5a共占0.25％),以及2个新基因型6变异体.所产生的5个BSP曲线图均凸显1993年至2000年这一时间段,中国的HCV感染数量呈现指数增长,随后则陡然下降.结论证实了HUV在中国多样性的流行现状,反映了HCV不断变化的基因型流行模式;1993年至2000年期间由于“单采血浆”事件的影响,HCV在中国的感染数量经历了一段“指数”增长期.
목적 탐토중국병형간염환자중HCV적기인형분포류형,병진행HCV재중국류행병학역사분석,진이연구HCV재중국적분자류행병학화진화동역학특정.방법 종423례병형간염환자적혈청중제취HCV RNA병진행cDNA반전록,확증E1화NS5B량개기인구적핵감산서렬,배제불합격급불괄합분석적병독주서렬,재대합격서렬진행계통발육분석(phylogenetic analysis).운용BEAST연건중적Bayesian MCMC (Markov Chain Monte Carlo)산법래진행Coalescence분석,통과중구BSPs(Bayesian skyline plots)도회소HCV적류행병학역사.결과 HCV분리주적기인분형여하:공포괄6개기인형,12개기인아형(1b:65.9％,6a:17.1％,2a:7.4％,3a:3.6％,3b:3.3％,6e:0.76％,1a,1c,2b,2f,4d이급5a공점0.25％),이급2개신기인형6변이체.소산생적5개BSP곡선도균철현1993년지2000년저일시간단,중국적HCV감염수량정현지수증장,수후칙두연하강.결론 증실료HUV재중국다양성적류행현상,반영료HCV불단변화적기인형류행모식;1993년지2000년기간유우“단채혈장”사건적영향,HCV재중국적감염수량경력료일단“지수”증장기.
Objective To determine the constantlychanging HCV genotype distribution patterns in China and investigate the HCV epidemic history and evolutionary dynamics.Methods HCV RNA from 423 HCV patients was extracted using Qiagen Viral RNA extraction kit,cDNA was transcripted using RevertAid First Strand cDNA Synthesis kit,and both E1 and NS5B sequences were amplified using RT-PCR followed by fluorescent sequencing.The sequences were analyzed using phylogenetic trees.The evolutionary analyses were performed using the Bayesian MCMC (Markov Chain Monte Carlo) algorithm implemented in the BEAST software and the BSP was reconstructed to form a retrospective picture of the epidemic history of HCV.Results Six HCV genotypes and 12 subtypes were identified (1b:65.9％,6a:17.1％,2a:7.4％,3a:3.6％,3b:3.3％,6e:0.76％,1a,1c,2b,2f,4d 5a:0.25％),in addition to two novel HCV-6 variants.A period from 1993 to 2000 was highlighted by the five BSPs,during which rapid HCV growth appeared followed by an abrupt slowing.Conclusions A great diversity in HCV was detected,which reflected a constantly changing pattern of HCV genotypes in China over time.They concurrently highlighted fast HCV growth in 1993-2000.This is a period during which HCV infection become explosive in China because of an officially encouraged plasma campaign in the1990s.