昆明市EV71病毒分离株3D蛋白酶序列特征、病毒增殖特性分析

发布时间：2018-07-23 07:42

【摘要】：目的比较昆明市住院与门诊病例来源的10株EV71病毒分离株3D蛋白酶基因核苷酸及编码氨基酸序列的差异性与病毒毒力之间的关系。方法对分离自昆明市的10株EV71病毒3D蛋白酶基因核苷酸测序,利用Bioedit及Mega 6软件对测序结果进行序列比对及遗传特性分析。选取6株(住院病例分离株、门诊病例分离株各3株)绘制病毒增殖动力学曲线,取其中2株(住院及门诊病例来源各1株)病毒接种至Vero细胞进行空斑形成试验,比较不同病毒株形成的空斑形态大小。结果对10株EV71分离病毒株的3D蛋白酶基因进行核苷酸测序并构建进化树,5株住院病例分离株间遗传距离较近,在进化树上单独成为一簇;住院病例病毒分离株与门诊病例病毒分离株的氨基酸序列在140、197、299位点存在差异;住院病例病毒株在较短时间达到增殖高峰,空斑试验中形成的蚀斑直径大于门诊病例分离株。结论不同来源的EV71病毒分离株3D蛋白酶基因核苷酸序列及编码氨基酸序列存在差异,该差异与EV71病毒的增殖特性及毒力具有一定相关性。
[Abstract]:Objective to compare the relationship between nucleotide sequence and coding amino acid sequence of 3D protease gene and virulence of 10 EV71 virus isolates in Kunming. Methods the nucleotide sequence of 3D protease gene of 10 strains of EV71 virus isolated from Kunming was sequenced, and the sequence alignment and genetic characteristics were analyzed by Bioedit and Mega6 software. Six strains (3 strains in hospital and 3 strains in outpatient department) were selected to draw the dynamic curve of virus proliferation, and 2 strains (1 strain from inpatient and 1 strain from outpatient) were inoculated into Vero cells for plaque formation test. The plaque size of different virus strains was compared. Results the 3D protease genes of 10 EV71 isolates were sequenced and 5 inpatient isolates were constructed. The amino acid sequence of inpatient virus isolate and outpatient virus isolate were different at the 140197299 locus, the inpatient virus strain reached the peak of proliferation in a short time, and the plaque diameter in plaque test was larger than that in outpatient case isolate. Conclusion the nucleotide sequence and coding amino acid sequence of 3D protease gene of EV71 virus isolates from different sources are different, which is related to the proliferation and virulence of EV71 virus.
【作者单位】：中国医学科学院医学生物学研究所北京协和医学院;
【基金】：中央级公益性科研院所基本科研业务费-“高等级生物安全P4实验室培训体系的建立”(No.2016ZX330058) 云南省创新团队-“中国医学科学院医学生物学研究所手足口病病原学与疫苗研究省创新团队”(No.2016HC009)
【分类号】：R373

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