A one-step reverse-transcription recombinase aided PCR assay for the rapid and sensitive detection of human enteroviruses  

作　　者：Xiuli Sun Huanhuan Lu Yanqing Tie Mengchuan Zhao Ruiqing Zhang Zhenlu Sun Guohao Fan Fengyu Li Fengyu Tian Yaxin Hu Mengyi Zhang Xinxin Shen Xuejun Ma Zhishan Feng 

机构地区：[1]North China University of Science and Technology,Tangshan 063210,China [2]Hebei General Hospital,Shijiazhuang 050051,China [3]NHC Key Laboratory of Medical Virology and Viral Diseases,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China [4]National Laboratory for Poliomyelitis,National Institute for Viral Disease Control and Prevention of the Chinese Center for Disease Control and Prevention,Beijing 102206,China [5]Hebei Key Laboratory of Molecular Medicine,Shijiazhuang 050051,China [6]Hebei Clinical Research Center for Laboratory Medicine,Shijiazhuang 050051,China [7]Department of Laboratory Medicine,Children's Hospital of Hebei Province,Shijiazhuang 050031,China [8]Yantai Center for Disease Control and Prevention,Yantai 264003,China

出　　处：《Biosafety and Health》2023年第2期126-131,共6页生物安全与健康（英文）

基　　金：This work was supported by the National Key R&D Program of China(2021YFC2301102);National Natural Science Foundation of China(82202593);the Central Guidance on Local Science and Technology Development Fund of Hebei Province(216Z7713G).

摘　　要：Human enteroviruses(HEVs)include many different types that cause a wide range of diseases,and an effective method of genus-level identification has therefore significant clinical implications.However,quantitative real-time reverse transcription polymerase chain reaction(qRT-PCR),the gold-standard method,still has shortfalls in diagnostic sensitivity and timeliness.Here we established a one-step real-time reverse-transcription recombinase-aided PCR assay(RT-RAP)to detect HEV fragment within an hour.The RT-RAP assay showed a detection limit of 5 copies/μL using recombinant plasmids and was extensively verified using 15 HEV strains.Among 15 types of HEV(species A-C),the sensitivity of RT-RAP was approximately 2-8 folds lower than that of the qRT-PCR in 9 types,and no-cross reaction with other viruses was observed.RT-RAP was further applied to analyze CSF and fecal specimens;the clinical performance demonstrated that the RT-RAP and the commercial qRT-PCR kit provided consistent results.These results indicated that RT-RAP assay may be a promising approach for rapid and sensitive detection of HEV.

关 键 词：Human enteroviruses Nucleic acid detection RT-RAP assay 

分 类 号：R446.5[医药卫生—诊断学]