RD、KMB-17及Vero细胞对肠道病毒分离培养敏感性的比较
Susceptibility comparison of enterovirus isolation and culture using RD,KMB-17 and Vero cells
刘煜菡 1张名 2许丹菡 2郭伟 2冯昌增 2徐丽兰 2马绍辉2
作者信息
- 1. 中国医学科学院 北京协和医学院医学生物学研究所 云南省重大传染病疫苗研发重点实验室,云南昆明 650118;自贡市疾病预防控制中心,四川自贡 643000
- 2. 中国医学科学院 北京协和医学院医学生物学研究所 云南省重大传染病疫苗研发重点实验室,云南昆明 650118
- 折叠
摘要
目的 比较人横纹肌肉瘤细胞RD、人胚肺二倍体细胞KMB-17和非洲绿猴肾细胞Vero对肠道病毒(enterovirus,EV)分离培养的敏感性,以期为后续EV各基因型毒株的分离提供实验依据.方法 收集2019年昆明市某医院确诊手足口病(hand-foot-mouth disease,HFMD)患者粪便样本60份,分别采用RD、KMB-17和Vero细胞分离培养EV,并对阳性样本进行RT-PCR检测、测序及NCBI BLAST比对,鉴定EV基因型.结果 共分离获得112株EV,分别属于11 种基因型.其中 RD细胞分离出 26株Echo-11,6株Echo-6和 Echo-30,2株CV-A4和 CV-A10,1 株CV-A6、CV-A 16和 Echo-18;KMB-17细胞分离出42株Echo-11,3株Echo-6和2株Echo-30;Vero细胞分离出 6株CV-B1,5株CV-B3,4株Echo-11,3株CV-A16和1株CV-B2.结论 RD细胞对大部分EV均敏感,KMB-17和Vero细胞分别对Echo和CV-B最敏感.
Abstract
Objective To compare the susceptibility of human rhabdomyosarcoma RD cells,human embryonic lung diploid KMB-17 cells and Vero cells to enterovirus(EV)isolation and culture,in order to provide experimental basis for subsequent isolation of EV genotypes.Methods Sixty stool samples of patients with hand-foot-mouth disease(HFMD)diagnosed in a hospital in Kunming in 2019 were collected,and EV strains were isolated and cultured with RD,KMB-17 and Vero cells respectively.The positive samples were detected by RT-PCR,sequenced and blasted by using NCBI BLAST software to identify EV genotypes.Results A total of 112 EV strains were isolated,belonging to 11 genotypes respectively.Among them,26 strains of Echo-11,6 strains of Echo-6 and Echo-30,2 strains of CV-A4 and CV-A10,1 strain of CV-A6,CV-A16and Echo-18 were isolated from RD cells;42 strains of Echo-11,3 strains of Echo-6 and 2 strains of Echo-30 were isolated from KMB-17cells;6 strains ofCV-B1,5 strains ofCV-B3,4 strains of Echo-11,3 strains of CV-A16 and 1 strain ofCV-B2 were isolated from Vero cells.Conclusion RD cells were susceptible to most EV genotypes,KMB-17 and Vero cells were the most susceptible to Echo virus and CV-B virus,respectively.
关键词
肠道病毒/细胞敏感性/Vero细胞/RD细胞/KMB-17细胞Key words
Enterovirus(EV)/Cell susceptibility/Vero cells/RD cells/KMB-17 cells引用本文复制引用
基金项目
云南省科技计划项目(202202AA100016)
云南省科技计划项目(202002AA100009)
云南省创新团队项目(202105AE160020)
云南省基础研究专项(20230-1AT070367)
出版年
2024
NETL
NSTL
万方数据