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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\u2010Cas12a\u2010fluorescence assay. The RPA\u2010Cas12a\u2010fluorescence system can specifically detect target gene levels as low as 0.5 copies of HBoV1 plasmid DNA per microliter within 40\u2009min at 37\u00b0C without the need for sophisticated instruments. The method also demonstrates excellent specificity without cross\u2010reactivity to non\u2010target 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\u2010Cas12a\u2010fluorescence assay, shows promising potential for early on\u2010site diagnosis of HBoV1 infection in the fields of public health and health care. The established RPA\u2010Cas12a\u2010fluorescence assay is rapid and reliable method for human bocavirus 1 detection. The RPA\u2010Cas12a\u2010fluorescence assay can be completed within 40\u2009min with robust specificity and sensitivity of 0.5 copies\/\u03bcl.<\/jats:p>","DOI":"10.1002\/ame2.12298","type":"journal-article","created":{"date-parts":[[2023,2,16]],"date-time":"2023-02-16T07:40:17Z","timestamp":1676533217000},"page":"179-188","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Development of <scp>RPA\u2010Cas12a<\/scp>\u2010fluorescence assay for rapid and reliable detection of human bocavirus 1"],"prefix":"10.1002","volume":"7","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4612-4351","authenticated-orcid":false,"given":"Weidong","family":"Qian","sequence":"first","affiliation":[{"name":"School of Food and Biological Engineering Shaanxi University of Science and Technology  Xi'an P. R. 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