论文部分内容阅读
目的对2009年中国云南省昆明地区柯萨奇病毒B5(Coxsackievirus group B5,CVB5)的部分VP1基因序列进行分析,以了解CVB5的遗传特性。方法收集2009年中国云南省昆明地区临床诊断为无菌性脑炎患儿的200份粪便标本,设计针对CVB的通用引物,利用半巢式RT-PCR扩增部分VP1基因。PCR产物直接测序后,采用Omiga和Mega 5.05等生物学软件进行核苷酸、氨基酸序列及系统进化分析。结果共分离出7株CVB5,其部分VP1基因之间的核苷酸和氨基酸序列同源性均较高,分别为92.7%～98.5%和93.6%～100.0%;与中国其他不同地区参考株的核苷酸序列同源性为80.1%～99.1%,其中与中国山东株98388-SD-CHN-1998同源性较低;与中国其他不同地区参考株的氨基酸序列同源性为84.4%～100.0%,其中与中国河南株CoxB5-Henan-2010和中国山东株98388-SD-CHN-1998之间同源性最低;与其他不同国家参考株比较,其核苷酸和氨基酸序列同源性分别在77.5%～89.1%和92.1%～96.3%之间;与原型株Faulkner的核苷酸和氨基酸序列同源性分别为79.6%～80.9%和92.1%～95.4%。基因系统进化分析显示,分离的7株CVB5属于相对独立的1个分枝,中国流行株除98388-SD-CHN-1998外,构成1个分枝。结论 2009年中国云南省昆明地区CVB5流行株属于相同基因型。
Objective To analyze the partial VP1 gene sequences of Coxsackievirus group B5 (CVB5) in Kunming, Yunnan Province, China in 2009 to understand the genetic characteristics of CVB5. Methods A total of 200 stool specimens from children with aseptic encephalitis in Kunming, Yunnan Province, China, were collected in 2009 and the universal primers for CVB were designed. The partial VP1 gene was amplified by semi-nested RT-PCR. After direct sequencing of the PCR products, biological software such as Omiga and Mega 5.05 were used for nucleotide, amino acid sequence and phylogenetic analysis. Results Seven strains of CVB5 were isolated. The nucleotide and amino acid sequences of some VP1 genes were highly homologous, ranging from 92.7% to 98.5% and 93.6% to 100.0%, respectively. Compared with other reference strains in China The nucleotide sequence homology was 80.1% -99.1%, which was lower than 98388-SD-CHN-1998 in Shandong China. The amino acid sequence homology was 84.4% -100.0 with reference strains in other regions of China %, Which has the lowest homology with CoxB5-Henan-2010 and China’s Shandong strain 98388-SD-CHN-1998. Compared with other reference strains in other countries, the nucleotide and amino acid sequence homologies were 77.5% -89.1% and 92.1% -96.3%, respectively. The nucleotide and amino acid sequences of Faulkner were 79.6% -80.9% and 92.1% -95.4%, respectively. Gene phylogenetic analysis showed that seven isolates of CVB5 belonged to a relatively independent branch, while the Chinese endemic strains except for 98388-SD-CHN-1998 constituted one branch. Conclusion In 2009, the epidemic strains of CVB5 in Kunming, Yunnan Province of China belonged to the same genotype.