浙江省2002年2例柯萨奇B_5病毒VP1区基因特性分析

Genetic characterization of the VP1 gene of the two strains of Coxsackie B_5 virus isolated from Zhejiang province in 2002

目的:研究2002年浙江省病毒性脑膜脑炎病原CoxB5病毒VP1区的基因特征.方法:用Hep-2和RD两种细胞对患者脑脊液和粪便标本进行病毒分离,提取病毒RNA,再用RT-PCR扩增病毒VP1区基因片断,对纯化产物进行核苷酸序列测定,采用DNAMAN和Bioedit软件进行分析处理.结果:两株CoxB5病毒的VP1区核苷酸长度均为 849 bp,二者核苷酸同源性为 98.7%,氨基酸同源性达 100%.结论:浙江省两株CoxB5病毒为同一性状毒株,它们的亲缘关系与侵犯中枢神经系统的CoxB5的原型株AF-114383CoxB5(Swend-16-1998)最为接近.但这两株CoxB5毒株与欧美国家相比,核苷酸同源性仅为 79.3%～81.5%,氨基酸同源性为 96.82～97.53%,核苷酸序列有了 18.5%～20.7% 的变异,与国外报道的CoxB5株之间存在一定的差异.这两株病毒与安徽明光市2001年从无菌性脑膜脑炎病人粪便中分离到的两株CoxB5病毒MG-18-2001和MG-39-2001在同一小分枝上,核苷酸同源性达 98.5%～98.9%,氨基酸同源性达 99.29%～99.65%.

Objective:To analyse the genetic characterization of the VP1 gene of Coxsackie B 5 virus causing meningitis isolated from Zhejiang province in 2002. Methods: The specimens of the cerebrospinal fluid (CSF) and feces were inoculated in Hep-2 and RD cell lines and two strains of Coxsackie B 5 virus were isolated. Viral RNA was extracted and the VP1 gene was amplified by RT-PCR. The sequence of the VP1 gene was determined and compared by DNAMAN and Bioedit software. Results: The VP1 region of the two strains has 849 bp and the genetic and protein homogeneity was 98.7% and 100% respectively. Conclusion: The two strains of Coxsackie B 5 virus showed the same characterization and had the nearest relationship with the prototype of AF-114383 Cox B 5(Swend-16-1998) which mostly infected human central nervous system. But the genetic and protein homogeneity was 79.3%～81.5% and 96.82%～97.53% respectively when they were compared with the Coxsackie B 5 virus isolated from the west and the variation was obvious. The genetic and protein homogeneity was 98.5%～98.9% and 99.29%～99.65% respectively when they were compared with the Coxsackie B 5 virus MG-18-2001 and MG-39-2001 isolated from the meningitis patients′ feces samples in Mingguang city of Anhui province in 2001.So these four strains were on the same branch of the phylogenetic tree.[