{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T19:28:29Z","timestamp":1792092509197,"version":"4.4.0"},"reference-count":33,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2025,5,14]],"date-time":"2025-05-14T00:00:00Z","timestamp":1747180800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,5,14]],"date-time":"2025-05-14T00:00:00Z","timestamp":1747180800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82273758"],"award-info":[{"award-number":["82273758"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100017957","name":"Health Commission of Guizhou Province","doi-asserted-by":"publisher","award":["gzwkj2024-055"],"award-info":[{"award-number":["gzwkj2024-055"]}],"id":[{"id":"10.13039\/100017957","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Disease Control and Prevention Administration","award":["Guo Jikong Zong Ren Han [2024]122"],"award-info":[{"award-number":["Guo Jikong Zong Ren Han [2024]122"]}]},{"name":"Foundation of Key Laboratory of Microbiology and Parasitology of Education Department of Guizhou","award":["QJJ [2022]019"],"award-info":[{"award-number":["QJJ [2022]019"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2025,5,14]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              Brucellosis is caused by members of the\n              <jats:italic toggle=\"yes\">Brucella<\/jats:italic>\n              spp. and remains one of the world\u2019s major zoonotic diseases.\n              <jats:italic toggle=\"yes\">Brucella melitensis<\/jats:italic>\n              (\n              <jats:italic toggle=\"yes\">B. melitensis<\/jats:italic>\n              ) as the most contagious\n              <jats:italic toggle=\"yes\">Brucella<\/jats:italic>\n              species cannot be ignored as an essential source of infection for brucellosis, especially in countries\/regions dominated by animal husbandry. Thus, the identification of the\n              <jats:italic toggle=\"yes\">Brucella<\/jats:italic>\n              genus and\n              <jats:italic toggle=\"yes\">B. melitensis<\/jats:italic>\n              is crucial for rapid diagnosis of brucellosis to control disease transmission and clinical treatment. Here, we developed the CRISPR\/Cas12b nuclease combined with a multiple cross displacement amplification (MCDA) assay (CRISPR-MCDA) for highly specific and sensitive detection of\n              <jats:italic toggle=\"yes\">Brucella<\/jats:italic>\n              genus and\n              <jats:italic toggle=\"yes\">B. melitensis<\/jats:italic>\n              in clinical applications. Two sets of specific primers were designed targeting the specific gene of\n              <jats:italic toggle=\"yes\">Brucella<\/jats:italic>\n              genus (\n              <jats:italic toggle=\"yes\">Bcsp31<\/jats:italic>\n              ) and\n              <jats:italic toggle=\"yes\">B. melitensis<\/jats:italic>\n              (\n              <jats:italic toggle=\"yes\">BMEII0466<\/jats:italic>\n              ), respectively. The CRISPR-MCDA assay showed high specificity and sensitivity in 28 non-\n              <jats:italic toggle=\"yes\">Brucella<\/jats:italic>\n              isolates and 64 clinical samples. The detection limit of CRISPR-MCDA assay was 2 copies\/\u03bcL in the plasmid dilution template, and the whole detection process took within 90 minutes with nanoparticle-based lateral flow biosensor (LFB) to validate experimental results. Taken together, the CRISPR-MCDA-LFB assay is a visual, sensitive, and highly specific detection technique that can be used as an attractive potential identification tool for\n              <jats:italic toggle=\"yes\">Brucella<\/jats:italic>\n              genus and\n              <jats:italic toggle=\"yes\">B. melitensis<\/jats:italic>\n              .\n            <\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>\n                The prevention and control of Brucellosis urgently require rapid and accurate diagnostic methods. This work validates a new method for the simultaneous detection of\n                <jats:italic toggle=\"yes\">Brucella<\/jats:italic>\n                genus and\n                <jats:italic toggle=\"yes\">B. melitensis<\/jats:italic>\n                . The method can effectively reduce the chances of contamination and provides a more rapid, sensitive, and specific on-site detection of\n                <jats:italic toggle=\"yes\">Brucella<\/jats:italic>\n                . It also offers a solution for the rapid screening of Brucellosis in resource-limited environments, which is crucial for effective disease prevention and control. This technology can also be widely applied to the rapid detection of other pathogens beyond\n                <jats:italic toggle=\"yes\">Brucella<\/jats:italic>\n                .\n              <\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/jcm.01532-24","type":"journal-article","created":{"date-parts":[[2025,4,11]],"date-time":"2025-04-11T13:02:11Z","timestamp":1744376531000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":12,"title":["Ultrasensitive and highly specific detection of the\n            <i>Brucella<\/i>\n            genus and\n            <i>B<\/i>\n            .\n            <i>melitensis<\/i>\n            by CRISPR\/Cas12b\u2010multiple cross displacement amplification technique"],"prefix":"10.1128","volume":"63","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-2206-688X","authenticated-orcid":false,"given":"Sha","family":"Mao","sequence":"first","affiliation":[{"name":"School of Public Health, the key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University","place":["Guiyang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4677-9470","authenticated-orcid":true,"given":"Xinggui","family":"Yang","sequence":"additional","affiliation":[{"name":"Guizhou Center for Disease Control and Prevention","place":["Guiyang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yue","family":"Wang","sequence":"additional","affiliation":[{"name":"Guizhou Center for Disease Control and Prevention","place":["Guiyang, 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