摘要
目的了解宁夏手足口病(HFMD)患者中柯萨奇病毒A14型(CVA14)分离株的基因特征。方法收集2013年实时荧光PCR法检测非EV71和非CVA16肠道病毒核酸阳性标本280份,用RD和Hep-2细胞进行肠道病毒分离培养,分离到的毒株采用RT-PCR法扩增其VP1区基因并进行核苷酸序列测定,测序结果利用BLAST进行型别鉴定。对鉴定为CVA14的毒株进行VP1区基因同源性分析和进化分析。结果从280份标本中共分离到肠道病毒36株,其中有2株鉴定为CVA14。2株CVA14宁夏分离株相互间核苷酸同源性为99.4%,氨基酸同源性为99.3%;与原型株G14的核苷酸同源性为82.3%~82.5%,氨基酸同源性为96.6%。系统进化显示,2株CVA14宁夏株与国内其他省份CVA14毒株处于同一分支。结论引起手足口病的病原除了优势病毒株EV71和CVA16外,同时还存在着CVA14等一些较少见、易忽略的肠道病毒共同循环的现象,因此应不断加强HFMD监测,以全面了解HFMD的流行趋势和抗原变异。
Objective To study the genetic characteristics of CVA14 strains isolated from the cases with hand,foot and mouth disease( HFMD) in Ningxia in 2013. Methods 280 specimens which identified as non- EV71,non- CVA16 entero- virus by real-time PCR were collected and cultured by using RD and Hep- 2 cells,and the VP1 gene of isolated stains was amplified by reverse transcriptase PCR( RT- PCR) with degenerate primers and sequenced. Sequences were compared with the database of Gen Bank by the BLAST algorithm to identify the virus genotypes. All the CVA14 strains identified were performed with homologous comparison and phylogenetic analysis. Results 36 virus strains were isolated from 280 clinical specimens,two strains were identified as CVAl4. The homologies of nucleotide and amino acid among Ningxia CVA14 strains were respectively 99. 4% and 99. 3%. The nucleotide homogeneity of Ningxia CVA14 strains with the G14 strains were 82. 3%- 82. 5%; the amino acid homogeneity were 96. 6%. Phylogenetic tree analysis revealed that Ningxia CVA14 strains and other province CVA14 strains in the same clusters. Conclusion In addition to EV71 and CVA16,other entero- virus serotypes such as CVA14 also could cause HFMD. The surveillance of HFMD should continue to be strengthened.
出处
《宁夏医学杂志》
CAS
2016年第6期517-519,共3页
Ningxia Medical Journal
基金
宁夏科研支撑资助项目(2013ZYS119)
关键词
手足口病
柯萨奇病毒A14
VP1区域
序列分析
Hand foot and mouth disease
Coxsackie virus A14
VP1 region
Sequence analysis
