{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T22:30:38Z","timestamp":1792103438046,"version":"4.4.0"},"reference-count":43,"publisher":"American Society for Microbiology","issue":"10","license":[{"start":{"date-parts":[[2025,10,7]],"date-time":"2025-10-07T00:00:00Z","timestamp":1759795200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,10,7]],"date-time":"2025-10-07T00:00:00Z","timestamp":1759795200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2025,10,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Brucellosis, a zoonotic disease caused by\n                      <jats:italic toggle=\"yes\">Brucella<\/jats:italic>\n                      species, presents significant public health challenges due to its complex diagnosis and the limited availability of rapid detection methods. To address these challenges, we developed a novel detection method that integrates recombinase polymerase amplification (RPA) with the CRISPR\/Cas12a system, enabling dual readout through fluorescence (FL) and lateral flow strip (LFS) detection. The RPA-CRISPR\/Cas12a-FL assay demonstrated an impressive detection limit of 1 copy\/\u03bcL, which is 10 times more sensitive than quantitative polymerase chain reaction, while the RPA-CRISPR\/Cas12a-LFS method achieved a detection limit of 10 copies\/\u03bcL, comparable to nested PCR. Specificity testing confirmed the robustness of the assay, as it produced strong signals exclusively for\n                      <jats:italic toggle=\"yes\">Brucella<\/jats:italic>\n                      without cross-reactivity with other bacterial species. Clinical validation using serum samples from 24 confirmed brucellosis patients and six healthy controls demonstrated a 100% concordance with serological results, underscoring the reliability of this method for clinical applications. This assay provides a rapid, sensitive, and specific tool for\n                      <jats:italic toggle=\"yes\">Brucella<\/jats:italic>\n                      detection, suitable for both laboratory and field settings, and holds significant potential for enhancing the diagnosis and control of brucellosis.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>Brucellosis is a significant zoonotic disease, and rapid and accurate diagnosis is crucial for its treatment and control. To address this need, we developed a novel detection method that combines recombinant enzyme polymerase amplification with a CRISPR\/Cas12a system, achieving dual readout through fluorescence and lateral flow strips. The test demonstrates excellent sensitivity and specificity, with clinical validation confirming complete concordance with serological results. This approach offers a fast, reliable, and field-deployable solution for brucellosis diagnosis, significantly enhancing disease management and public health outcomes.<\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.00998-25","type":"journal-article","created":{"date-parts":[[2025,8,27]],"date-time":"2025-08-27T13:00:29Z","timestamp":1756299629000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":17,"title":["Development of a rapid and sensitive RPA-CRISPR\/Cas12a-based assay for the detection of\n                    <i>Brucella melitensis<\/i>"],"prefix":"10.1128","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-0864-6558","authenticated-orcid":true,"given":"Yu","family":"Shen","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00dr1cn74","id-type":"ROR","asserted-by":"publisher"}],"name":"Hangzhou Center for Disease Control and 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