人B组轮状病毒荧光定量RT-PCR检测方法的建立  

Real-Time Reverse Transcriptase Polymerase Chain Reaction for Detection and Quantification of Human Group B Rotavirus

作　　者：毛彤瑶 李金松[2] 王萌璇 刘诗琪 孙晓曼[2] 庞立丽[2] 张佳艳[3] 裴银辉 段招军[2] 李丹地[2] MAO Tongyao;LI Jinsong;WANG Mengxuan;LIU Shiqi;SUN Xiaoman;PANG Lili;ZHANG Jiayan;PEI Yinhui;DUAN Zhaojun;LI Dandi(School of Basic Scicences,North China University of Sciences and Technology,Tangshan 063210,China;Key Laboratory of Medical Viruses and Viral Diseases,National Health Commission,National Institute for Viral Disease Control and Prevention,Chinese Center for Diseases Control and Prevention,Beijing 102206,China;Lushan College,University of Science and Technology,Liuzhou 545616,China)

机构地区：[1]华北理工大学基础医学院,唐山063210 [2]国家卫生健康委员会医学病毒和病毒病重点实验室,中国疾病预防控制中心病毒病预防控制所,北京102206 [3]广西科技大学鹿山学院,柳州545616

出　　处：《病毒学报》2020年第6期1092-1100,共9页Chinese Journal of Virology

基　　金：国家科技重大专项(项目号:2018ZX10711-001),题目:艾滋病和病毒性肝炎等重大传染病防治;国家自然科学基金(项目号:8160813),题目:P[6]基因型轮状病毒跨物种传播的分子机制研究。

摘　　要：人B组轮状病毒曾在我国发生过散发和暴发流行,近年来在世界多个国家也均有报道。为建立一种灵敏和特异的人B组轮状病毒荧光定量RT-PCR检测方法,本研究针对人B组轮状病毒NSP3片段保守区设计和筛选特异性引物和探针,优化反应体系和反应条件,并对该方法的灵敏性、特异性、稳定性进行评价。结果显示,该方法针对人B组轮状病毒检出限可以达到6.28拷贝/μL;与多种常见和罕见的人腹泻病毒没有交叉反应;批内和批间等重复性试验变异系数均小于5%。本研究建立的人B组轮状病毒荧光定量RT-PCR检测方法灵敏度高、特异性强、稳定性好、检出率高,可作为一种新的技术手段应用于人B组轮状病毒的快速筛查与诊断。Human group B rotavirus has been sporadic and outbreak in China,and has been reported in many countries in the world in recent years.We wished to establish a sensitive and specific fluorescent quantitative reverse transcription-polymerase chain reaction(RT-PCR)for detection of human group B rotaviruses(RVB).We designed and screened specific primers and probes according to the conserved region of the NSP3 fragment of RVB.We optimized the reaction and evaluated the sensitivity,specificity and stability of the method.The limit of detection of RVB was 6.28 copies/μL,and there was no cross-reaction with various common or rare viruses that cause human diarrhea.The coefficient of variation within and between assays was<5%.The fluorescent quantitative RT-PCR for RVB established in this study had high sensitivity,specificity,stability and detection.Our method could be used as new technology for the rapid screening and diagnosis of RVB.

关 键 词：人B组轮状病毒 荧光定量RT-PCR 疾病防控 

分 类 号：R373.2[医药卫生—病原生物学]