论文部分内容阅读
目的研究绍兴市2012—2013年手足口病患者的肠道病毒71型(EV 71)病毒分离株的基因特征。方法采集手足口病患者粪便、疱疹液和咽拭子标本,采用逆转录聚合酶链反应(RT-PCR)特异性扩增进行EV 71鉴定,阳性的样本进行病毒分离,对10株分离株分A、B、C、D和E五段进行分段PCR,测序后并拼接成全长,以EV 71 A、B和C各基因型的参考毒株和以往中国EV 71的分离毒株为参考株,基于核苷酸序列和氨基酸序列对VP1区分别进行同源性分析和系统发生树构建。结果 RT-PCR结果证实分离的10株病毒均为EV 71,与C4a亚型亲缘关系最近,它们之间核苷酸和氨基酸同源性分别为95.06%~97.87%和93.60%~98.99%;与亲缘关系最远的参考毒株Cox A16的核苷酸和氨基酸同源性分别为61.17%~62.74%和68.00%~71.38%。在系统发生树上,这10株EV 71均在C基因型中,与属C4a亚型的病毒株属同一分支。结论绍兴地区手足口病患者中分离的EV 71毒株属C基因型的C4a亚型进化分支。
Objective To study the genetic characteristics of Enterovirus 71 (EV 71) isolated from patients with hand-foot-mouth disease in Shaoxing during 2012-2013. Methods Feces, herpes fluid and throat swab specimens were collected from patients with hand-foot-mouth disease. EV71 was identified by reverse transcription-polymerase chain reaction (RT-PCR) specific amplification. Positive samples were isolated by virus. Ten isolates A, B, C, D and E five segments of segmented PCR, sequenced and spliced into full length, with EV 71 A, B and C genotype reference strains and past China EV 71 isolates as reference strains The homology analysis and phylogenetic tree construction of the VP1 region were carried out based on the nucleotide sequence and the amino acid sequence respectively. Results The RT-PCR results showed that all of the 10 isolates were EV 71, which had the closest genetic relationship with C4a subtypes. The nucleotide and amino acid homologies between them were 95.06% -97.87% and 93.60% -98.99%, respectively. The nucleotide and amino acid homology of the reference strain Cox A16 with the closest genetic relationship were 61.17% -62.74% and 68.00% -71.38%, respectively. In the phylogenetic tree, these 10 strains of EV 71 are all in the C genotype, belonging to the same branch as the strain belonging to the genus C4a. Conclusion The EV 71 strain isolated from patients with HFMD in Shaoxing belongs to the C4a subtype evolutionary branch of C genotype.