{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T06:34:50Z","timestamp":1773297290998,"version":"3.50.1"},"reference-count":30,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,3,2]],"date-time":"2022-03-02T00:00:00Z","timestamp":1646179200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:p><jats:italic>Nocardia cyriacigeorgica<\/jats:italic> has gradually become a common pathogen in clinical microbial infections. Identification of <jats:italic>Nocardia<\/jats:italic> at the species level is essential to assess the susceptibility and pathogenicity of antimicrobials. However, there is no suitable method for rapid and accurate laboratory detection of <jats:italic>N. cyriacigeorgica<\/jats:italic>. In this study, we combined PCR amplification with the CRISPR-Cas12a system to establish a novel detection platform, named CRISPR-PCR, and applied it to the detection of <jats:italic>N. cyriacigeorgica<\/jats:italic> in clinical samples. The Cas12a protein exhibited collateral cleavage activity following CRISPR RNA binding to specific targets, then indiscriminately cleaved nearby single-stranded DNA, and this was evaluated for diagnostic nucleic acid detection by measuring the fluorescence signal using a fluorescence reader. The assay takes only 2 h, including DNA extraction for 20 min, nucleic acid pre-amplification for 70 min, and fluorescence detection for 20 min. The limit of detection for <jats:italic>N. cyriacigeorgica<\/jats:italic> was 10<jats:sup>-3<\/jats:sup> ng and the specificity was 100%. Thus, the <jats:italic>N. cyriacigeorgica<\/jats:italic> CRISPR-PCR assay is a rapid and specific method for detecting <jats:italic>N. cyriacigeorgica<\/jats:italic>, and the CRISPR-PCR fluorescence detection platform has great potential for detection of other pathogens.<\/jats:p>","DOI":"10.3389\/fcimb.2022.835213","type":"journal-article","created":{"date-parts":[[2022,3,2]],"date-time":"2022-03-02T07:56:20Z","timestamp":1646207780000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":10,"title":["A CRISPR-Cas12a-Assisted Fluorescence Platform for Rapid and Accurate Detection of Nocardia cyriacigeorgica"],"prefix":"10.3389","volume":"12","author":[{"given":"Xueping","family":"Liu","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaotong","family":"Qiu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuai","family":"Xu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yanlin","family":"Che","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lichao","family":"Han","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yutong","family":"Kang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuan","family":"Yue","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shenglin","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fang","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhenjun","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2022,3,2]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaf5573","article-title":"C2c2 Is a Single-Component Programmable RNA-Guided RNA-Targeting CRISPR Effector","volume":"353","author":"Abudayyeh","year":"2016","journal-title":"Science"},{"key":"B2","doi-asserted-by":"publisher","first-page":"4895","DOI":"10.1128\/JCM.43.9.4895-4897.2005","article-title":"Brain Abscess Caused by Nocardia Cyriacigeorgica in a Patient With Human Immunodeficiency Virus Infection","volume":"43","author":"Barnaud","year":"2005","journal-title":"J. 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