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目的了解江苏省2012-2014年柯萨奇病毒A组16型(coxsackievirus A16,CA16)流行毒株基因型概况。方法收集江苏省2012-2014年儿童手足口病标本并送样,进行病毒分离培养获取相应毒株,用CA16特异性引物通过反转录聚合酶链反应技术进行VP1区基因片段扩增和测序,利用生物信息学软件对序列进行分析,并与CA16参比株序列进行同源性比较并构建基因进化树。结果分离得到82株CA16毒株,测序结果显示全部属于基因亚型B1,VP1基因分析显示其中9株毒株序列的基因亚型为B1a,另外73株毒株序列的基因亚型为B1b。CA16分离株VP1区核苷酸和氨基酸同源性分别为87.8%~100.0%和97.5%~100.0%;与CA16国际标准株G10核苷酸和氨基酸同源性分别为75.2%~78.2%和90.5%~91.9%。结论江苏省2012-2014年分离获得CA16毒株属于基因亚型B1,以B1a和B1b两个分支共同进化和流行,其中又以B1b为优势亚型。
Objective To understand the genotypes of coxsackievirus A16 (CA16) strains in Jiangsu province from 2012 to 2014. Methods Samples of hand, foot and mouth disease in children from 2012 to 2014 in Jiangsu Province were collected and sent for virus isolation and culture to obtain corresponding strains. The VP1 gene fragment was amplified and sequenced by reverse transcription-polymerase chain reaction using CA16-specific primers. Bioinformatics software was used to analyze the sequences and to compare homology with CA16 reference strains and construct phylogenetic tree. Results 82 strains of CA16 were isolated and sequenced. All the isolates belonged to genotype B1. The analysis of VP1 gene showed that the genotypes of nine strains were B1a and the other 73 were B1b. The nucleotide and amino acid homologies of VP1 of CA16 were 87.8% -100.0% and 97.5% -100.0%, respectively. The nucleotide and amino acid homologies of G16 to CA16 were 75.2% -78.2% and 90.5%, respectively % ~ 91.9%. Conclusion The CA16 strain isolated from Jiangsu province from 2012 to 2014 belonged to genotype B1, which co-evolved and popularized in both B1a and B1b branches. Among them, B1b was the dominant subtype.