论文部分内容阅读
目的了解北京市西城区2015年手足口病(HFMD)患儿肠道病毒71型(EV 71)分离株VP1区基因特征,分析其变异情况。方法收集北京市西城区2015年HFMD病原学检测分型鉴定为EV 71的阳性标本,对部分标本进行病毒分离、培养,并通过实时荧光PCR鉴定阳性培养物。EV 71阳性培养物提取病毒核酸,对其VP1区全长序列进行PCR扩增和测序。从Genbank中选取EV 71不同基因型别参考序列,利用生物信息学软件进行同源性和系统进化分析。结果共分离到6株EV 71(Genbank登录号为KU376383-KU376388),其VP1区核苷酸和氨基酸序列同源性分别为93.00%~99.00%和98.00%~100.00%,与参考序列VP1区核苷酸和氨基酸序列同源性分别为81.00%~97.00%和93.00%~100.00%。进化分析显示,6株EV 71均为C4a基因亚型。结论 2015年北京市西城区EV 71流行株属于C4a亚型,未出现明显变异。
Objective To investigate the characteristics of VP1 gene of enterovirus 71 (EV 71) isolated from children with hand-foot-mouth disease (HFMD) in Xicheng District of Beijing in 2015 and analyze the variation. Methods The positive samples of EV71 were identified by pathogen detection of HFMD in 2015 in Xicheng District of Beijing. Viruses were isolated and cultured in some samples. Positive cultures were identified by real-time fluorescence PCR. The EV71 positive culture was used to extract the viral nucleic acid and amplify and sequence the full-length VP1 region of the virus. EV71 genotype reference sequences were selected from GenBank, and homology and phylogenetic analysis were performed using bioinformatics software. Results A total of 6 strains of EV71 (GenBank accession No. KU376383-KU376388) were isolated. The nucleotide and amino acid sequences of VP1 were 93.00% -99.00% and 98.00% -100.00%, respectively. The nucleotide and amino acid sequence homologies were 81.00% ~ 97.00% and 93.00% ~ 100.00%, respectively. Phylogenetic analysis showed that all of the six EV71 strains were subtypes C4a. Conclusion EV 71 epidemic strains in Xicheng District of Beijing belong to C4a subtype in 2015, with no obvious variation.