基于杂交链式(HCR)反应的甲型流感病毒检测技术研究（英文）  

Hybridization Chain Reaction-based Assay for The Detection of Influenza A Virus

作　　者：孙明璇[1,2] 赵荣涛 李杨[2] 孔文[2] 杨益 郭旭东[2] 刘婉莹 宋宏彬 张志红[1] 郝荣章[2] SUN Ming-Xuan;ZHAO Rong-Tao;LI Yang;KONG Wen;YANG Yi;GUO Xu-Dong;LIU Wan-Ying;SONG Hong-Bin;ZHANG Zhi-Hong;HAO Rong-Zhang(Deportment of Environmental Health, School of Public Health, Shanxi Medical University, Taiyuan 030001, China;Institute of Disease Control and Prevention, PLA, Beijing 100071, China)

机构地区：[1]山西医科大学公共卫生学院环境卫生教研室,太原030001 [2]中国人民解放军疾病预防控制所,北京100071

出　　处：《生物化学与生物物理进展》2018年第6期644-652,共9页Progress In Biochemistry and Biophysics

基　　金：supported by grants from National Science and Technology Major Project of China(2018ZX10711001003002,2015AA020929);Beijing Nova Program(Z141107001814071)~~

摘　　要：甲型流感病毒的现场快速检测对于流感的及时有效防控具有重要意义.本研究基于杂交链式(HCR)反应,利用GO对荧光基团的猝灭作用及共同实现了对甲型流感病毒的快速检测.当目标序列存在时,可引发HCR反应,使短链DNA形成长链,保护FAM荧光基团不被猝灭,从而实现目标物的检测.实验结果表明,该方法在10～40 nmol/L范围内荧光强度与目标检测物浓度表现出了良好的线性关系,检测范围为5～100 nmol/L.这种HCR等温扩增检测技术具有较好的样本检测能力,具有等温、无酶、反应体系简单、操作步骤简便等优点,表现出良好的现场检测应用前景.Rapid point-of-care（POC） detection of influenza A virus is critical for timely and effective flu prevention and control. In this study, we developed a hybridization chain reaction（HCR）-based assay, through coupling HCR reaction with fluorescence quenching by graphene oxide（GO）, for the rapid detection of influenza A virus. The target is detected as it triggers HCR, which results in the extension of short DNA chains, protecting the 6-carboxy-fluorescein amidite（FAM） group from GO quenching. The results demonstrate that it could specifically recognize target nucleic acid fragment of influenza A virus from other pathogen, and even from single-base mismatched oligonucleotides. A good linear correlation between fluorescence intensity and the target concentrations ranging from 10 to 40 nmol/L was achieved, with a detection limit of 5 nmol/L. Its detection performance was verified on nasopharyngeal swab samples, which is the first clinical application of HCR-based influenza assay. This HCR-based method shows many advantages including the enzyme-free amplification of nucleic acids, simple reaction system and convenient protocols, suggesting its availability for POC detection

关 键 词：甲型流感 杂交链式反应 氧化石墨烯 现场检测 无酶检测 

分 类 号：R440[医药卫生—诊断学]