Development of RPA-Cas12a-fluorescence assay for rapid and reliable detection of human bocavirus 1

作     者：Weidong Qian Xuefei Wang Ting Wang Jie Huang Qian Zhang Yongdong Li Si Chen Weidong Qian;Xuefei Wang;Ting Wang;Jie Huang;Qian Zhang;Yongdong Li;Si Chen

作者机构：School of Food and Biological EngineeringShaanxi University of Science and TechnologyXi'anP.R.China Department of DermatologyHuazhong University of Science and Technology Union Shenzhen HospitalShenzhenP.R.China Ningbo Municipal Center for Disease Control and PreventionNingboP.R.China Medical College of Shenzhen UniversityShenzhenP.R.China 

出 版 物：《Animal Models and Experimental Medicine》 (动物模型与实验医学（英文）)

年 卷 期：2024年第7卷第2期

页      面：179-188页

核心收录：

学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 1001[医学-基础医学(可授医学、理学学位)] 100103[医学-病原生物学] 10[医学] 

基　　金：Natural Science Foundation of China Grant/Award Number:81973531 Science and Technology Plan Project of Xi’an Grant/Award Number:22GXFW0007 Shenzhen Science and Technology Innovation Commission Grant/Award Number:20200812211704001 Medical Scientific Research Foundation of Guangdong Province Grant/Award Number:A2019502 Nanshan District Science and Technology Plan Project Grant/Award Number:NS2022022 Scientific Research Program Funded by Shaanxi Provincial Education Department Grant/Award Number:22JC010 

主　　题：CRISPR-Cas12a detection human bocavirus 1 on-site diagnosis recombinase polymerase amplification 

摘      要：Human bocavirus(HBoV)1 is considered an important pathogen that mainly affects infants aged 6–24 months,but preventing viral transmission in resource-limited regions through rapid and affordable on-site diagnosis of individuals with early infection of HBoV1 remains somewhat ***,we present a novel faster,lower cost,reliable method for the detection of HBoV1,which integrates a recombinase polymerase amplification(RPA)assay with the CRISPR/Cas12a system,designated the RPA-Cas12a-fluorescence *** RPA-Cas12a-fluorescence system can specifically detect target gene levels as low as 0.5 copies of HBoV1 plasmid DNA per microliter within 40 min at 37℃without the need for sophisticated *** method also demonstrates excellent specificity without cross-reactivity to non-target ***,the method was appraised using 28 clinical samples,and displayed high accuracy with positive and negative predictive agreement of 90.9%and 100%,***,our proposed rapid and sensitive HBoV1 detection method,the RPA-Cas12a-fluorescence assay,shows promising potential for early on-site diagnosis of HBoV1 infection in the fields of public health and health *** established RPA-Cas12a-fluorescence assay is rapid and reliable method for human bocavirus 1 *** RPA-Cas12a-fluorescence assay can be completed within 40 min with robust specificity and sensitivity of 0.5 copies/μl.