{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T18:40:14Z","timestamp":1791312014620,"version":"4.3.1"},"reference-count":28,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,2,25]],"date-time":"2021-02-25T00:00:00Z","timestamp":1614211200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002855","name":"Ministry of Science and Technology of the People's Republic of China","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002855","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002855","name":"Ministry of Science and Technology of the People's Republic of China","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002855","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100016308","name":"Wuxi Health and Family Planning Commission","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100016308","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:p><jats:italic>Vibrio cholerae<\/jats:italic> and <jats:italic>Vibrio parahaemolyticus<\/jats:italic> are common diarrheal pathogens of great public health concern. A multiplex TaqMan-based real-time PCR assay was developed on the BD MAX platform; this assay can simultaneously detect and differentiate <jats:italic>V. cholerae<\/jats:italic> and <jats:italic>V. parahaemolyticus<\/jats:italic> directly from human fecal specimens. The assay includes two reactions. One reaction, BDM-VC, targets the gene <jats:italic>ompW<\/jats:italic>, the cholera toxin (CT) coding gene <jats:italic>ctxA<\/jats:italic>, the O1 serogroup specific gene <jats:italic>rfbN<\/jats:italic>, and the O139 serogroup specific gene <jats:italic>wbfR<\/jats:italic> of <jats:italic>V. cholerae<\/jats:italic>. The other, BDM-VP, targets the gene <jats:italic>toxR<\/jats:italic> and the toxin coding genes <jats:italic>tdh<\/jats:italic> and <jats:italic>trh<\/jats:italic> of <jats:italic>V. parahaemolyticus<\/jats:italic>. In addition, each reaction contains a sample process control. When evaluated with spiked stool samples, the detection limit of the BD MAX assay was 195\u2013780 CFU\/ml for <jats:italic>V. cholerae<\/jats:italic> and 46\u2013184 CFU\/ml for <jats:italic>V.\u00a0parahaemolyticus<\/jats:italic>, and the amplification efficiency of all genes was between 95 and 115%. The assay showed 100% analytical specificity, using 63 isolates. The BD MAX assay was evaluated for its performance compared with conventional real-time PCR after automated DNA extraction steps, using 164 retrospective stool samples. The overall percent agreement between the BD MAX assay and real-time PCR was \u2265 98.8%; the positive percent agreement was 85.7% for <jats:italic>ompW<\/jats:italic>, 100% for <jats:italic>toxR<\/jats:italic>\/<jats:italic>tdh<\/jats:italic>, and lower (66.7%) for <jats:italic>trh<\/jats:italic> because of a false negative. This is the first report to evaluate the usage of the BD MAX open system in detection and differentiation of <jats:italic>V. cholerae<\/jats:italic> and <jats:italic>V. parahaemolyticus<\/jats:italic> directly from human samples.<\/jats:p>","DOI":"10.3389\/fcimb.2021.639473","type":"journal-article","created":{"date-parts":[[2021,2,25]],"date-time":"2021-02-25T08:33:51Z","timestamp":1614242031000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["Development of a Rapid and Fully Automated Multiplex Real-Time PCR Assay for Identification and Differentiation of Vibrio cholerae and Vibrio parahaemolyticus on the BD MAX Platform"],"prefix":"10.3389","volume":"11","author":[{"given":"Zhenpeng","family":"Li","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongxia","family":"Guan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"He","family":"Gao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weihong","family":"Feng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baowei","family":"Diao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongqun","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Biao","family":"Kan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingyun","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,2,25]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0003832","article-title":"Updated Global Burden of Cholera in Endemic Countries","volume":"9","author":"Ali","year":"2015","journal-title":"PLoS Negl. 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