人A组G2P[4]型轮状病毒的分离培养和鉴定  

Isolation, culture and identification of a group A human rotavirus G2P[4] strain

作　　者：高升辉 王明雯 李利利[2] 李丹地[2] 宋敬东 王雅璐 毛彤瑶 吴建军 李金松[2] 段招军[2] Gao Shenghui;Wang Mingwen;Li Lili;Li Dandi;Song Jingdong;Wang Yalu;Mao Tongyao;Wu Jianjun;Li Jinsong;Duan Zhaojun(School of Public Health,Gansu University of Chinese Medicine,Lanzhou 730099,China;National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China)

机构地区：[1]甘肃中医药大学公共卫生学院,兰州730099 [2]中国疾病预防控制中心病毒病预防控制所,北京102206

出　　处：《国际病毒学杂志》2022年第2期93-97,共5页International Journal of Virology

摘　　要：目的对我国轮状病毒A组G2P[4]流行株进行分离培养和鉴定。方法采用轮状病毒抗原胶体金法筛查56份病毒性腹泻患儿粪便标本,对阳性样本VP7和VP4基因测序确定其G和P基因型别,并接种至MA-104细胞,持续传代。运用SDS-PAGE电泳、实时荧光定量PCR、间接免疫荧光、嗜斑实验和电镜等技术进行鉴定。结果检出轮状病毒阳性样本46份,测序确定G2P[4]型2株,细胞培养盲传5代,嗜斑纯化后G2P[4]型毒株RNA PAGE电泳图呈DS-1株轮状病毒的(4:2:3:2)形态,电镜下呈车轮状形态,轮廓清晰,病毒滴度随时间变化逐步上升。结论成功从粪便样本中分离和培养出G2P[4]型A组轮状病毒。Objective Isolation,culture and identification of the prevalent strain of rotavirus G2P[4]in China.Methods The stool samples of 56 children with viral diarrhea were tested for rotavirus antigen by colloidal gold method.The positive samples were inoculated into MA-104 cells and maintained continuous propagation.The strains were identified by SDS-PAGE,real-time PCR,indirect immunofluorescence,plaque test and electron microscopy.Results A total of 46 rotavirus positive samples were detected.Sequencing identified 2 strains of G2P[4].After blind passage of 5 generations and plaque purification,the PAGE electrophoregram of the RNA of G2P[4]strain showed patterns similar to DS-1 strain of rotavirus(4:2:3:2).The virus showed a clear morphology of wheel under electron microscope.The growth kinetic curve of the virus increased gradually with time.Conclusions Successful isolation and culture the strain of group A rotavirus G2P[4]from fecal samples.

关 键 词：轮状病毒 分离培养 PAGE电泳 电镜 生长曲线 

分 类 号：R373.2[医药卫生—病原生物学]