2009年中国云南昆明地区柯萨奇病毒A组16型遗传特性分析

Genetic characters of coxsackievirus A16 strain in 2009 in Kunming Region,Yunnan Province,China

目的对2009年中国云南昆明地区柯萨奇病毒A组16型(Coxsackievirus A16,CoxA16)毒株的部分VP1区进行分析,了解CoxA16的遗传特性。方法设计针对肠道病毒A群的通用引物,采用半巢式RT-PCR扩增目的片段,对PCR产物测序;使用Omiga和Mega4.1等生物学软件进行核苷酸、氨基酸序列比对及病毒的系统进化分析。结果200份2009年昆明市儿童医院临床诊断为手足口病或并发脑炎疑似肠道病毒感染的患者粪便标本中有52份为CoxA16感染,其中15份来自脑炎患者。52株CoxA16病毒部分VP1区核苷酸和氨基酸序列同源性均较高,核苷酸序列同源性为98.5%～100%,氨基酸序列同源性为98.8%～100%;与shzh00-2、shzh99-48和107/Toyama/1990相比较,核苷酸序列同源性为92.3%～93.8%,氨基酸序列同源性为95.3%～96.8%;与国际标准株G10相比较,核苷酸序列同源性为78.5%～80.3%,氨基酸序列同源性为93.2%～94.5%;与其他13个参考株的核苷酸序列同源性为96.6%～99.5%,氨基酸序列同源性为98.3%～100%。基因系统进化分析显示,52株CoxA16均属于B基因型的一个分支。结论 2009年中国云南昆明地区CoxA16株属于B2亚型。

Objective To analyze the partial VP1 domain of coxsackievirus A 16 （CoxA 16） strain isolated in 2009 in Kunming Region, Yunnan Province, China so as to investigate the genetic characters of the strain. Methods Universal primers specific to enterovirus group A were designed, with which the target gene fragment was amplified by semi-nested RT-PCR and sequenced. The deduced nucleotide and amino acid sequences of amplified gene were analyzed by biological software including Omiga and Mega4. 1, based on which a phylogenetic tree was plotted. Results A total of 200 fecal samples of suspected patients with hand-foot-and-mouth disease or complicated encephalitis, caused by enterovirns infection, were collected from Kunming Children＇s Hospital in 2009, of which 52 were positive for CoxA16, including 15 ones from patients with encephalitis. The homologies of nucleotides and amino acids of partial VP1 domains of the 52 CoxA16 strains were 98. 5% - 100% and 98. 8% - 100% respectively. However, the homologies of nucleotides and amino acids of CoxA16 strain were 92. 3% - 93. 8% and 95. 3% - 96. 8% to those of shzh00-2, shzh99-48 and 107/ Toyama/1990 strains, 78. 5% - 80. 3% and 93. 2% - 94. 5% to those of international standard strain G10, and 96. 6% - 99. 5% and 98. 3% - 100% to those of other 13 reference strains, respectively. Phylogenetic analysis proved that all the 52 CoxA16 strains belonged to a branch of genotype B. Conclusion The CoxA16 strains isolated in 2009 in Kunming Region, Yunnan Province, China were of subtype B2.