2019-2020年湖北部分地区G9P[8]型人轮状病毒基因特征  

Genetic characteristics of human rotavirus with the G9P[8]genotype in Part Area of Hubei Province from 2019 to 2020

作　　者：杨立君 陈月 米文琴 唐一通[2] 范久波[2] 王媛 廖诗英[2] 陈焕春[1] 何启盖[1] 江云波[1] YANG Li-jun;CHEN Yue;MI Wen-qin;TANG Yi-tong;FAN Jiu-bo;WANG Yuan;LIAO Shi-ying;CHEN Huan-chun;HE Qi-gai;JIANG Yun-bo(State Key Laboratory of Agricultural Microbiology,College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China;Department of Medicine,Hubei University of Arts and Sciences,Wuhan 430070,China)

机构地区：[1]华中农业大学动物医学院农业微生物学国家重点实验室,武汉430070 [2]湖北文理学院医学部,武汉430070

出　　处：《中国人兽共患病学报》2022年第4期359-365,共7页Chinese Journal of Zoonoses

基　　金：中央高校基本科研业务费专项基金资助(No.2662019YJ005);国家现代产业技术体系(No.CARS-35)联合资助。

摘　　要：目的对人A组轮状病毒进行检测及分离鉴定,并研究其各基因片段的遗传进化关系。方法2019-2020年对湖北武汉市和襄阳市临床腹泻病人的粪便样品进行采集,共319份。设计特异性轮状病毒VP6基因引物,RT-PCR检测轮状病毒的感染情况。将阳性样品接种于MA104细胞进行轮状病毒的分离。RT-PCR特异性扩增VP6基因和特异性间接免疫荧光对其进行病毒鉴定及病毒增殖检测;并进一步通过RT-PCR扩增轮状病毒的11个基因片段,在线工具Rota C V2.0对测序结果进行分型分析。Mega软件对其全基因组序列进行遗传进化分析。结果轮状病毒感染阳性标本共69份,阳性率为21.63%。成功分离获得11株人轮状病毒,主要衣壳蛋白VP7和VP4基因型均为G9P[8]型。其中3株轮状病毒归属于类Wa株毒株,基因型图谱为G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1。8株毒株在Wa-like的基因型中具有DS-1-like的NSP4为E2基因型特征。基因型图谱为G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E2-H1。结论G9P[8]型人轮状病毒毒株在2019-2020年湖北部分地区占主导趋势,且其NSP4基因以E2基因型为主要流行形式。Human rotavirus group A was detected,isolated and identified,and the genetic evolutionary relationships among its gene fragments were studied.A total of 319 diarrhea-related samples from hospitals in Wuhan and Xiangyang,Hubei province,were collected from 2019 to 2020.Specific primers for the rotavirus VP6 gene were designed,and rotavirus infection was detected with RT-PCR.The positive samples were inoculated into MA104 cells for rotavirus isolation.The virus was identified through amplification of the VP6 gene and specific indirect immunofluorescence.Then 11 rotavirus gene fragments were amplified by RT-PCR,and the sequencing results were typed with Rota C v2.0.The whole genome sequence was analyzed in mega software.A total of 69 samples were found to be positive for rotavirus,and the positivity rate was 21.63%.Eleven human rotavirus strains were successfully isolated.The main capsid protein VP7 and VP4 genotypes of all strains were G9P[8].Three rotavirus strains were wa like strains,and the genotype constellation was G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1.Among the wa like genotypes,eight strains had the E2 genotype characteris tic for NSP4 of DS-1-like strains.The genotype constellation was G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E2-H1.G9P[8]human rotavirus strains remained dominant in some areas of Hubei province from 2019 to 2020,and the NSP4 gene of the epidemic strains was of the E2 genotype.The results lay a foundation for further study of the characteristics of rotavirus epidemic strains and prevention.

关 键 词：轮状病毒 分离鉴定 基因型 遗传进化分析 

分 类 号：R373.2[医药卫生—病原生物学]