急性呼吸道感染儿童中WU多瘤病毒基因组序列特征分析  

Genomic Characterization of WU Polyomavirus Identified from Pediatric Patients with Acute Respiratory Infections in Beijing,China

作　　者：赵林清[1] 钱渊[1] 朱汝南[1] 邓洁[1] 王芳[1] 孙宇[1] 丁雅馨[1] 张尼娜[1] 

机构地区：[1]首都儿科研究所病毒研究室,北京100020

出　　处：《病毒学报》2010年第6期447-452,共6页Chinese Journal of Virology

基　　金：国家自然科学基金(30872153);北京市科技新星计划(2006A63)

摘　　要：为了解从北京地区急性呼吸道感染儿童中发现的WU多瘤病毒的基因组编码特征,并对其进行基因序列多样性分析,应用针对基因组5'端非编码区、衣壳蛋白VP1、VP2编码基因以及LTAg编码基因的引物对,从已确证为WU病毒阳性的来自北京地区急性呼吸道感染儿童的编号为BJF5276的临床标本中经聚合酶链反应扩增得到预期的基因片段,直接测序后将序列拼接得到全基因组序列,进而推导其基因组编码特征;随后从其它21例已确证为WU多瘤病毒阳性的急性呼吸道感染儿童标本中扩增得到衣壳蛋白VP2编码区基因,进行基因序列测定以及基因序列多样性分析。得到了WU病毒BJF5276全基因组序列。序列分析结果显示WU病毒BJF5276基因组序列全长为5229bp,共有5个主要的CDS(Coding domain sequences),分别编码衣壳蛋白VP2、VP3、VP1,并以其互补序列为模板,编码STAg和LTAg;所得到的22例VP2蛋白编码区基因序列同源性比较结果显示病毒VP2基因编码区序列与GenBank中已有的64个序列之间同源性很高;Mega4.0NJ进化树(Neighbor-joiningtree)分析显示这22个VP2基因序列分属于不同的基因进化簇,其中20个序列属于进化簇I中的Ia,另外2个序列属于进化簇III,其中的一个序列在IIIb基因进化簇中,另外一个序列独立成簇,不属于现有的IIIa或IIIb,暂时将其命名为IIIc。本研究结果提示北京地区的WU病毒具有多瘤病毒科的基因组编码特性;序列非常保守,有分属于不同基因进化簇的WU病毒在北京地区流行,与文献报道的以Ib流行为主所不同的是北京地区的WU病毒以Ia为主,且有新的基因进化簇出现。To characterize the genomic sequence and arrangement of WU polyomavirus（WU virus） identified in clinical specimens collected from children with acute respiratory infections in Beijing,China,the sequences of capsid proteins VP1,VP2,and the large tumor antigen（LTAg）,as well as the 5＇-terminal sequence of WU virus,were amplified from the clinical specimen with ID number of BJF5276 which was determined as WU virus positive by PCR amplification.The PCR amplicons were sequenced,and genomic sequence analysis was performed by using the software DNAStar.In addition,VP2 coding-region sequences were amplified from other 21 clinical specimens identified as WU virus positive to investigate the gene diversity of WU virus.The genomic sequence of WU virus BJF5276 with accession number of HQ218321 in GenBank was 5,229 base pairs in length with 3 major coding domain sequences（CDS） sited on one strand coding for capsid proteins VP2,VP3 and VP1,and two CDS sited on the complementary strand coding for small tumor antigen（STAg） and LTAg;These 22 VP2 CDS sequences including 5 sequences submitted to GenBank were compared with 64 corresponding sequences downloaded from GenBank by MegAlign of DNAStar software,indicated that these sequences coming from children in Beijing shared high homology（over 98.8%） with those from GenBank.Phylogenetic analysis of these VP2 CDS by using Neighbor-joining（NJ） analyses with 2,000 bootstraps（Mega 4.0） showed that 20 sequences out of 22 belonged to clade Ia,and other 2 of them belonged to clade III,including 1 clustered in IIIa and 1 in a novel cluster proposed as IIIc.In conclusion,the genomic sequence of WU polyomavirus detected from clinical specimens from children in Beijing is closely related to other WU polyomaviruses in the feature of genomic coding region arrangement.Overall variation of VP2 CDS was very low,and there were different clades circulating in Beijing with a dominant clade Ia,which is different from dominated Ib circulating in other parts of the world report

关 键 词：WU多瘤病毒(WU病毒) 基因序列分析 急性呼吸道感染 儿童 

分 类 号：R373.1[医药卫生—病原生物学] R373.9[医药卫生—基础医学]