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

作　　者：刘煜菡 张名[1] 许丹菡 郭伟 冯昌增[1] 徐丽兰[1] 马绍辉[1] LIU Yuhan;ZHANG Ming;XU Danhan;GUO Wei;FENG Changzeng;XU Lilan;MA Shaohui(Institute of Medical Biology,Chinese Academy of Medical Sciences&Peking Union Medical College,Key Laboratory of Vaccine Research and Development for Major Infectious Diseases in Yunnan Province,Kunming 650118,Yunnan Province,China)

机构地区：[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 [2]自贡市疾病预防控制中心,四川自贡643000

出　　处：《中国生物制品学杂志》2024年第8期917-920,共4页Chinese Journal of Biologicals

基　　金：云南省科技计划项目(202202AA100016,202002AA100009);云南省创新团队项目(202105AE160020);云南省基础研究专项(20230-1AT070367).

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-A16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。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 1l 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-A16 and 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-17 cells;6 strains of CV-B1,5 strains of CV-B3,4 strains of Echo-11,3 strains of CV-A16 and 1 strain of CV-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 

分 类 号：R373.2[医药卫生—病原生物学]