{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T07:01:25Z","timestamp":1789974085899,"version":"4.0.1"},"reference-count":47,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,9,20]],"date-time":"2026-09-20T00:00:00Z","timestamp":1789862400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,20]],"date-time":"2026-09-20T00:00:00Z","timestamp":1789862400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Fish Diseases"],"abstract":"ABSTRACT<\/jats:title>\n \n Aquaculture associated\n Vibrio<\/jats:italic>\n species are important bacterial pathogens causing significant economic losses in aquaculture and zoonotic infections in humans. Rapid and accurate detection of\n Vibrio<\/jats:italic>\n species is therefore essential for diagnosis, surveillance and control. In this study, a multiplex quantitative PCR (qPCR) assay was developed for the simultaneous detection of\n Vibrio<\/jats:italic>\n spp. together with\n \n V. alginolyticus<\/jats:italic>\n <\/jats:styled-content>\n ,\n \n V. anguillarum<\/jats:italic>\n <\/jats:styled-content>\n ,\n \n V. harveyi<\/jats:italic>\n <\/jats:styled-content>\n ,\n \n V. ordalii<\/jats:italic>\n <\/jats:styled-content>\n ,\n \n V. parahaemolyticus<\/jats:italic>\n <\/jats:styled-content>\n ,\n \n V. vulnificus<\/jats:italic>\n <\/jats:styled-content>\n and\n \n V. cholerae<\/jats:italic>\n <\/jats:styled-content>\n . Species\u2010specific primers and probes targeting\n ompA, glutamate synthase, mreB, vohB, dnaJ, gyrB<\/jats:italic>\n and\n lolB<\/jats:italic>\n genes were designed, while the\n rpoA<\/jats:italic>\n gene was selected for genus level\n Vibrio<\/jats:italic>\n detection. The assay demonstrated 100% inclusivity for all target strains without cross reactivity against non\u2010target bacterial species. PCR efficiency\/inhibition analyses showed stable amplification dynamics across serial dilutions, with\n R<\/jats:italic>\n 2<\/jats:sup>\n values above 0.99, amplification efficiencies between 96.8% and 98.2%, and \u0394Cq values \u2264\u20090.13 for all targets. Limit of detection (LOD) studies performed using artificially contaminated matrices demonstrated a detection limit of 10\n 1<\/jats:sup>\n \u201310\n 0<\/jats:sup>\n \u2009CFU\/mL for all targets, with detection performance comparable to the culture\u2010based recovery method. Overall, the developed multiplex qPCR assay provides a rapid, sensitive and analytically reliable approach for simultaneous detection of multiple\n Vibrio<\/jats:italic>\n species associated with aquaculture and zoonotic infections.\n <\/jats:p>","DOI":"10.1111\/jfd.70289","type":"journal-article","created":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T06:13:15Z","timestamp":1789971195000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of a Multiplex Quantitative\n PCR<\/scp>\n Assay for Simultaneous Detection of Multiple\n Vibrio<\/i>\n Species in Aquaculture Samples"],"prefix":"10.1111","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9180-5768","authenticated-orcid":false,"given":"\u0130nci Ba\u015fak","family":"M\u00fc\u015ftak","sequence":"first","affiliation":[{"name":"Department of Microbiology, Faculty of Veterinary Medicine Ankara University Ankara T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hamit Kaan","family":"M\u00fc\u015ftak","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Veterinary Medicine Ankara University Ankara T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2386-6857","authenticated-orcid":false,"given":"Seyyide Sar\u0131\u00e7am","family":"\u0130nce","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Veterinary Medicine Ankara University Ankara T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-2297-5319","authenticated-orcid":false,"given":"Do\u011fukan","family":"U\u011fun","sequence":"additional","affiliation":[{"name":"Bioeksen R&D Technologies Incorporated Istanbul T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9853-565X","authenticated-orcid":false,"given":"G\u00f6rkem","family":"Cengiz","sequence":"additional","affiliation":[{"name":"Department of Food Hygiene and Technology, Faculty of Veterinary Medicine Ankara University Ankara T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8996-7028","authenticated-orcid":false,"given":"G\u00fcltekin","family":"\u00dcnal","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Veterinary Medicine Ankara University Ankara T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8230-4427","authenticated-orcid":false,"given":"Ebru","family":"Torun","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Veterinary Medicine Ankara University Ankara T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7245-1941","authenticated-orcid":false,"given":"Muammer","family":"G\u00f6nc\u00fco\u011flu","sequence":"additional","affiliation":[{"name":"Department of Food Hygiene and Technology, Faculty of Veterinary Medicine Ankara University Ankara T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,20]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"crossref","DOI":"10.3389\/fmicb.2021.617703","article-title":"Molecular Detection and Distribution of Six Medically Important Vibrio spp. in Selected Freshwater and Brackish Water Resources in Eastern Cape Province, South Africa","volume":"12","author":"Abioye O. 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X.Zou andJ.Li.2009. \u201cDetection ofVibrio anguillarumby PCR Amplification of the empA Gene.\u201d32:293\u2013296.","DOI":"10.1111\/j.1365-2761.2008.00984.x"},{"key":"e_1_2_11_46_1","doi-asserted-by":"publisher","DOI":"10.3390\/vetsci12040327"},{"key":"e_1_2_11_47_1","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.mcp.2011.02.002","article-title":"SYBR Green I\u2010Based Real\u2010Time PCR Targeting the rpoX Gene for Sensitive and Rapid Detection of \n Vibrio alginolyticus","volume":"25","author":"Zhao J. 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