人H组轮状病毒免疫荧光检测方法的建立  

Development of an Immunofluorescence Method for Detection of Human Rotavirus H

作　　者：庞立丽[1] 白爱英[2] 卿钰 张佳艳[3] 李伯阳 章青[1] 李丹地[1] 梁未丽[5] 阚飙[5] 段招军[1] PANG Lili;BAI Aiying;QING Yu;ZHANG Jiayan;LI Boyang;ZHANG Qing;LI Dandi;LIANG Weili;KAN Biao;DUAN Zhaojun(National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Jinan Center for Disease Control and Prevention,Jinan 250021,China;Lushan College,Gugangxi University of Science and Technology,Liuzhou 545000,China;College of Elementary Medicine,North China University of Science and Technology,Tangshan 063000,China;National Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China)

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京102206 [2]济南市疾病预防控制中心,济南250021 [3]广西科技大学鹿山学院,柳州545000 [4]华北理工大学基础医学院,唐山063000 [5]中国疾病预防控制中心传染病预防控制所,北京102206

出　　处：《病毒学报》2020年第6期1085-1091,共7页Chinese Journal of Virology

基　　金：“十三五”国家科技重大专项课题(项目号:2018ZX10301408-001),题目:急性病毒感染相关的胞内固有免疫生物标志物的发现与应用。

摘　　要：轮状病毒(Rotavirus,RV)是引起婴幼儿病毒性胃肠炎的主要病原体。目前,尚没有人H组轮状病毒(Rotavirus H,RVH)的免疫荧光(Immunofluorescence,IF)检测方法。本研究建立了检测人H组轮状病毒的免疫荧光方法,为研究轮状病毒感染以及与宿主免疫反应之间的作用机制提供技术支撑。本研究首先培养MA104恒河猴肾细胞,感染人H组轮状病毒J19毒株,对病毒进行培养;原核表达纯化病毒VP6蛋白,用纯化的VP6蛋白制备antiVP6抗体;以anti-VP6作为检测抗体,应用免疫荧光检测技术检测J19病毒在MA104细胞中的定位;并利用免疫荧光方法检测J19病毒的荧光灶形成单位(Fluorescence focus units,FFU),对病毒进行定量。本方法为进一步检测和研究人H组轮状病毒感染机制提供技术条件。Rotaviruses are the major cause of viral gastroenteritis in children,but there is no method of immunofluorescence(IF)detection for human rotavirus H(RVH).This study established an IF method for detection of RVH,providing technical support for the study of rotavirus infection and its interaction with host immune response.First,MA104 cells were cultured and infected with rotavirus strain J19.The VP6 protein of J19 was prokaryotically expressed and purified,and was used to immunize rabbits to produce antibody against VP6.The IF assay was established with anti-VP6 as detection antibody.Localization of J19 virus in MA104 cells was determined by IF assay.The quantity of J19 virus was evaluated by the fluorescence focus units method.The methods proposed in this study can contribute to the further study of the infection mechanism of RVH.

关 键 词：H组轮状病毒(RVH) 细胞培养 VP6蛋白 免疫荧光 

分 类 号：R373.2[医药卫生—病原生物学]