DOI: 10.1002/ame2.12298
Development of RPA-Cas12a-fluorescence assay for rapid and reliable detection of human bocavirus 1
Weidong Qian1
Xuefei Wang1
Ting Wang1
Jie Huang1
Qian Zhang2
Yongdong Li3
Si Chen4
1.School of Food and Biological Engineering,Shaanxi University of Science and Technology,Xi'an,P.R.China2.Department of Dermatology,Huazhong University of Science and Technology Union Shenzhen Hospital,Shenzhen,P.R.China3.Ningbo Municipal Center for Disease Control and Prevention,Ningbo,P.R.China4.Medical College of Shenzhen University,Shenzhen,P.R.China
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 re-gions 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 spe-cifically 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 detec-tion 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/μ l.
机标关键词：developmentfluorescencedetectionhumanrapidassaybocavirusreliable
论文发表日期：2024-04-28
在线出版日期：2025-08-15（本平台首次上网日期，不代表文献的发表时间）
页数：10( 179-188 )
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