Zhao Lin-Qing, Qian Yuan, Zhu Ru-Nan, Deng Jie, Wang Fang, Sun Yu, Ding Ya-Xin, Zhang Ni-Na
Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China.
Bing Du Xue Bao. 2010 Nov;26(6):447-52.
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 reported previously, and a novel clade IIIc was proposed.
为了鉴定从中国北京急性呼吸道感染儿童临床标本中分离出的WU多瘤病毒（WU病毒）的基因组序列及排列情况，从编号为BJF5276的临床标本中扩增衣壳蛋白VP1、VP2以及大T抗原（LTAg）的序列，还有WU病毒的5'端序列，该标本经PCR扩增确定为WU病毒阳性。对PCR扩增产物进行测序，并使用DNAStar软件进行基因组序列分析。此外，从另外21份鉴定为WU病毒阳性的临床标本中扩增VP2编码区序列，以研究WU病毒的基因多样性。GenBank中登录号为HQ218321的WU病毒BJF5276基因组序列长度为5229个碱基对，一条链上有3个主要编码域序列（CDS），编码衣壳蛋白VP2、VP3和VP1，互补链上有2个CDS，编码小T抗原（STAg）和LTAg；使用DNAStar软件的MegAlign将这22条VP2 CDS序列（包括提交至GenBank的5条序列）与从GenBank下载的64条相应序列进行比较，结果表明来自北京儿童的这些序列与GenBank中的序列具有高度同源性（超过98.8%）。使用邻接法（NJ）并进行2000次自展分析（Mega 4.0）对这些VP2 CDS进行系统发育分析，结果显示22条序列中有20条属于Ia分支，另外2条属于III分支，其中1条聚在IIIa分支，1条在一个新提出的IIIc分支中。总之，从北京儿童临床标本中检测到的WU多瘤病毒基因组序列在基因组编码区排列特征上与其他WU多瘤病毒密切相关。VP2 CDS的总体变异非常低，北京有不同分支在传播，其中Ia分支占主导，这与之前报道的世界其他地区占主导的Ib分支不同，并且提出了一个新的IIIc分支。
