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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。
In order to understand the genomic coding characteristics of WU polyoma virus found in children with acute respiratory tract infection in Beijing and to analyze the diversity of its gene sequences, aiming at the genomic non-coding region of 5 ’, capsid protein VP1, VP2 encoding genes and LTAg encoding gene from the clinical validation of WU virus-positive children from Beijing acute respiratory tract number BJF5276 clinical samples amplified by polymerase chain reaction amplification of the expected gene fragments, the direct sequencing of the sequence splicing obtained Genomic sequence and then deduced its coding characteristics of the genome. Then, the VP2 coding region of the capsid protein was amplified from 21 other children confirmed to be WU polyoma positive by acute respiratory infection, and the gene sequences were determined and the gene sequences were diverse Sexual analysis. Got WU virus BJF5276 whole genome sequence. The results of sequence analysis showed that the genome of WU virus BJF5276 was 5229bp in length and contained 5 major CDS sequences encoding the capsid proteins VP2, VP3 and VP1, respectively, encoding the STAg and LTAg with their complementary sequences as templates. The results of homology analysis of the coding region of VP2 protein in 22 cases showed that the homology between the coding region of VP2 gene and 64 existing sequences in GenBank was high. Neighbor-joining tree of Mega4.0NJ, The analysis showed that the 22 VP2 gene sequences belong to different gene evolution clusters, among which 20 sequences belong to Ia in evolutionary cluster I, the other 2 sequences belong to evolution cluster III, one of them is in the IIIb gene evolution cluster, and the other The sequences are clustered independently and do not belong to the existing IIIa or IIIb, tentatively named IIIc. The results of this study suggest that the WU virus in Beijing has the polyomavirus family of genomic coding characteristics. The sequence is very conservative, and WU viruses that belong to different gene evolution clusters are endemic in Beijing, which is different from those reported in the literature WU virus in Beijing is mainly Ia, and a new gene cluster appears.