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目的通过测定和分析肠道病毒71型毒株VP1区序列,掌握了EV71的流行情况及种系进化关系,为手足口病毒所致疾病的预防和治疗提供支持和依据。方法利用RT-PCR的方法扩增EV71病毒VP1区核苷酸序列,测序后利用DNASTAR和Primer Premier 5.0进行序列比对分析,并用Mega 3.1软件建立检测样本VP1区序列与Gen Bank上EV71基因型参考毒株基因序列的系统发育树。结果随机选取的21株EV71分离株的核苷酸和氨基酸序列同源性分别为95.6%~100%和99.2%~100%;通过系统进化树分析显示,21株EV71均属于C4a基因亚型。结论 2016年杭州地区EV71毒株的基因亚型与当前国内外某些地区的分型结果一致,故它们有望成为疫苗候选株并作进一步研究。
OBJECTIVE: To determine the prevalence of EV71 and its phylogenetic relationship by detecting and analyzing the VP1 region of enterovirus 71 strain and provide support and evidence for preventing and treating hand-foot-mouth disease. Methods The nucleotide sequence of VP1 region of EV71 virus was amplified by RT-PCR. Sequence analysis was performed using DNASTAR and Primer Premier 5.0 after sequencing. Mega 3.1 software was used to establish the VP1 region of the test sample and the genotype of EV71 on Gen Bank Phylogenetic tree of strain gene sequences. Results The nucleotide and amino acid sequences of 21 EV71 isolates randomly selected were 95.6% -100% and 99.2% -100%, respectively. Phylogenetic tree analysis showed that 21 EV71 belonged to C4a genotypes. Conclusion The genotypes of EV71 strains in Hangzhou area are consistent with the typing results in some areas at home and abroad in 2016, so they are expected to become vaccine candidate strains for further study.