1株轮状病毒疫苗衍生株的分离和鉴定  

Isolation and identification of a vaccine-derived rotavirus strain

作　　者：何莲花 冯瑄[3] 陈燕 姜慧敏 李金松[2] 张少白[3] 段招军[2] He Lianhua;Feng Xuan;Chen Yan;Jiang Huimin;Li Jinsong;Zhang Shaobai;Duan Zhaojun(College of Public Health,Gansu University of Traditional Chinese Medicine,Lanzhou 730000,Gansu,China;National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Shaanxi Center for Disease Control and Prevention,Xi'an 710054,Shaanxi,China;School of Basic Medicine,North China University of Science and Technology,Tangshan 063200,Hebei,China)

机构地区：[1]甘肃中医药大学公共卫生学院,甘肃兰州730000 [2]中国疾病预防控制中心病毒病预防控制所,北京102206 [3]陕西省疾病预防控制中心,陕西西安710054 [4]华北理工大学基础医学院,河北唐山063200

出　　处：《中国疫苗和免疫》2023年第5期548-553,共6页Chinese Journal of Vaccines and Immunization

基　　金：国家重点研发计划(2021YFC2301000);陕西省创新能力支撑计划(2022PT-26)。

摘　　要：目的对1例免疫功能正常的急性胃肠炎(Acute gastroenteritis,AGE)成人患者感染的轮状病毒(Rotavirus,RV)疫苗衍生株(Vaccine-derived strain,VDS)进行鉴定和探究其AGE病因。方法采集与1名接种口服五价人-牛重配RV减毒活疫苗(Pentavalent oral live human-bovine reassortant rotavirus vaccine,RV5)儿童密切接触且患AGE的母亲的粪便标本,采用实时逆转录聚合酶链反应(Real-time reverse transcription-polymerase chain reaction,Real-time RT-PCR)、RT-PCR或PCR筛查RV等常见致AGE的病毒,采用MA104细胞对RV进行分离培养,通过RT-PCR扩增和测序获得全基因组,采用聚丙烯凝胶电泳进行双链RNA电泳带型鉴定,利用在线分型工具分型,进行Blastn和Blastp序列比对。结果从该AGE患者粪便标本中筛查出RV;细胞培养在盲传第二代时产生明显的细胞病变效应;该分离株经VP4/VP7基因测序显示为源于RV5的G4P[5]基因型RV5 VDS,且呈现A组RV独特的4∶2∶3∶2双链RNA电泳图谱;与RV5原型株相比,该分离株的NSP3节段核苷酸发生G283 T变异,推导氨基酸发生D95Y变异,其他节段的核苷酸和氨基酸序列相似性为100%。结论该成人AGE患者感染的RV5 VDS可能来自接种RV5的儿童,但导致该成人AGE的可能性较小;尚需加强监测以评估其公共卫生意义。Objective To identify a vaccine-derived strain(VDS)of rotavirus(RV)in an adult acute gastroenteritis(AGE)patient with normal immune function,and to explore AGE causes.Methods We collected fecal specimens from a mother with AGE,a close contact of her child who had received pentavalent oral live human-bovine reassortant rotavirus vaccine(RV5).We used real-time reverse transcription-polymerase chain reaction(real-time RT-PCR),RT-PCR,or PCR to screen for RV and other common viruses causing AGE.The RV was isolated and cultured using MA104 cell and its whole genome was obtained by RT-PCR amplification and sequence detection.The whole genome of RV was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then typed by online typing tools.Sequence analysis was conducted with Blastn and Blastp tools.Results An RV strain was detected from fecal specimens of the AGE patient.Obvious cytopathic effect occurred during the second generation of blind cell culture.The sequencing of VP4/VP7 genes showed that the RV isolate was a strain of G4P[5]genotype of RV5 VDS.The whole genome showed a unique 4∶2∶3∶2 double-stranded RNA electrophoretic map of group A RV.Compared with the prototype strain of RV5,a nucleic acid mutation of G283 T and a deduced amino acid mutation of D95Y occurred at the NSP3 segment of the isolate;similarities of nucleotide and amino acid sequences were 100%at other segments.Conclusions The RV5 VDS isolated from the AGE patient may have come from the child who received RV5,but her AGE was unlikely caused by the isolate.We should strengthen surveillance to assess the public health significance of RV5 VDS.

关 键 词：轮状病毒 轮状病毒疫苗 疫苗衍生株 急性胃肠炎 

分 类 号：R186[医药卫生—流行病学]