我国人群感染指环病毒的高通量测序分析  

Characterization of anelloviruses in the Chinese population using high-throughput sequencing

作　　者：程实[1,2] 范航[2] 孙强[2] 裴广倩[2] 安小平[2] 米志强[2] 张湘莉兰 黄勇[2] 童贻刚[1,2] 

机构地区：[1]安徽医科大学军事医学科学院微生物流行病研究所,安徽合肥230032 [2]军事医学科学院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京100071

出　　处：《中国病原生物学杂志》2016年第10期865-870,共6页Journal of Pathogen Biology

基　　金：艾滋病和病毒性肝炎等重大传染病防治科技重大专项"十二五"实施计划(No.2013ZX10004-605,2013ZX10004-217,2013ZX10004-607,2011ZX10004-001);国家高技术研究发展计划(863计划,2014AA021402,2012AA022-003,2015AA020108);国家自然科学基金项目(No.81572045)

摘　　要：目的分析我国人群感染指环病毒的多样性,为研究指环病毒科的感染与不同疾病发生以及宿主免疫状态之间的关系提供基础资料。方法从33份血液标本中提取病毒核酸,使用多重置换扩增技术富集指环病毒核酸,然后使用Ion Torrent PGM平台进行高通量测序,并进行生物信息学分析。结果共得到226个新的指环病毒序列,包括130条全序和96个ORF1。受检血液标本均检出包括TTV,TTMDV和TTMV等指环病毒;测序显示指环病毒中81.9%为TTV,48.5%的标本感染两个属以上的指环病毒。指环病毒ORF1至少81.13%发生变异,其中TTV属内也存在75.66%的变异,13.27%的序列在NCBI的核酸数据库中同源比对未成功。进化分析显示TTV基因3型又可分为3.1、3.2、3.3等3个亚型,以往命名的基因2型是基因3.3型的一个子分枝。新型TTV中51.9%属于基因3型。结论我国人群普遍感染指环病毒,以基因3型感染居多且基因组多样性高。Objective To characterize the diversity of anelloviruses in the Chinese population to provide basic information on the role of infection by the family Anelloviridae in various diseases and the immune status of different hosts.Methods Multiple displacement amplification was used as an initial anellovirus enrichment tool,followed by highthroughput sequencing.Afterwards,bioinformatic methods were used to analyze the sequencing results as well as the status of virus infection in blood samples. Results A total of 226 sequences from 33 individuals were determined to be from anelloviruses based on BLASTN or BLASTX analysis.Of these sequences,130 were complete genome sequences and the other 96 were ORF1sequences.At least one strain of anellovirus was found in sequenced samples from humans and 81.9% of the anellovirus strains were the Torque teno virus（TTV）.Of all of the samples,48.5% were infected with two or more genera of TTV,TTMDV,or TTMV.The three genera had up to 81.13% genomic variation among the nucleotide sequences of the ORF1 gene and 75.66% genomic variation among the genus Torque teno virus according to pairwise comparison.BLASTN using default parameters for megablast indicated that 13.27% of the sequences did not coincide with sequences in the NT nucleotide database of the National Center for Biotechnology Information（NCBI）.Phylogenetic analysis indicated that the lineages of genotype 3were separated into three subtypes,namely subtype 3.1,3.2,and 3.3,with the previously reported genotype 2as a branch of subtype 3.3.Fifty-one-point-nine percent of the sequenced TTV fell into genotype 3,suggesting that subtype 3viruses comprised the majority of anelloviruses infecting the Chinese population.The long evolutionary distance also reflected the high level of diversity of anelloviruses,and especially TTV,infecting the Chinese population. Conclusion Results revealed the high prevalence of anelloviruses in the Chinese population,indicating that high-throughput sequencing combined with multiple displacement ampl

关 键 词：指环病毒 多重置换扩增(MDA) 输血传播病毒(TTV) 进化 

分 类 号：R373.9[医药卫生—病原生物学]