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 

机构地区：[1]School of Food and Biological Engineering,Shaanxi University of Science and Technology,Xi'an,P.R.China [2]Department of Dermatology,Huazhong University of Science and Technology Union Shenzhen Hospital,Shenzhen,P.R.China [3]Ningbo Municipal Center for Disease Control and Prevention,Ningbo,P.R.China [4]Medical College of Shenzhen University,Shenzhen,P.R.China

出　　处：《Animal Models and Experimental Medicine》2024年第2期179-188,共10页动物模型与实验医学（英文）

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

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 challenging.Herein,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 assay.The 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 instruments.The method also demonstrates excellent specificity without cross-reactivity to non-target pathogens.Furthermore,the method was appraised using 28 clinical samples,and displayed high accuracy with positive and negative predictive agreement of 90.9%and 100%,respectively.Therefore,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 care.The established RPA-Cas12a-fluorescence assay is rapid and reliable method for human bocavirus 1 detection.The RPA-Cas12a-fluorescence assay can be completed within 40 min with robust specificity and sensitivity of 0.5 copies/μl.

关 键 词：CRISPR-Cas12a DETECTION human bocavirus 1 on-site diagnosis recombinase polymerase amplification 

分 类 号：R373[医药卫生—病原生物学]