{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T12:47:37Z","timestamp":1791377257266,"version":"4.3.2"},"reference-count":45,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,6,28]],"date-time":"2023-06-28T00:00:00Z","timestamp":1687910400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:p>Bovine pasteurellosis, caused by serogroups A, B, and E of <jats:italic>Pasteurella multocida<\/jats:italic> (Pm), is mainly manifested as bovine respiratory disease (BRD) and hemorrhagic septicemia (HS). The disease has caused a great economic loss for the cattle industry globally. Therefore, identifying the Pm serogroups is critical for optimal diagnosis and subsequent clinical treatment and even epidemiological studies. In this study, a one-step multiplex real-time PCR assay was established. Three pairs of specific primers were prepared to detect the highly conserved genomic regions of serogroups A (<jats:italic>HyaD<\/jats:italic>), B (<jats:italic>bcbD<\/jats:italic>), and E (<jats:italic>ecbJ<\/jats:italic>) of Pm, respectively. The results depicted that the method had no cross-reaction with other bovine pathogens (<jats:italic>Mannheimia hemolytica, Escherichia coli, Listeria monocytogenes, Staphylococcus aureus, Salmonella Dublin, Mycobacterium paratuberculosis<\/jats:italic>, infectious bovine rhinotracheitis virus, and <jats:italic>Mycoplasma bovis<\/jats:italic>). The linear range (10<jats:sup>7<\/jats:sup> to 10<jats:sup>2<\/jats:sup> copies\/\u03bcL) showed the R<jats:sup>2<\/jats:sup> values for serogroups A, B, and E of Pm as 0.9975, 0.9964, and 0.996, respectively. The multiplex real-time PCR efficiency was 90.30%, 90.72%, and 90.57% for CartA, CartB, and CartE, respectively. The sensitivity result showed that the serogroups A, B, and E of Pm could be detected to be as low as 10 copies\/\u03bcL. The repeatability result clarified that an intra-assay and an inter-assay coefficient of variation of serogroups A, B, and E of Pm was &amp;lt; 2%. For the clinical samples, the detection rate was higher than the OIE-recommended ordinary PCR. Overall, the established one-step multiplex real-time PCR assay may be a valuable tool for the rapid and early detection of the serogroups A, B, and E of Pm with high specificity and sensitivity.<\/jats:p>","DOI":"10.3389\/fvets.2023.1193162","type":"journal-article","created":{"date-parts":[[2023,6,28]],"date-time":"2023-06-28T09:04:21Z","timestamp":1687943061000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":12,"title":["Construction of a one-step multiplex real-time PCR assay for the detection of serogroups A, B, and E of Pasteurella multocida associated with bovine pasteurellosis"],"prefix":"10.3389","volume":"10","author":[{"given":"Haojie","family":"Wang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingxiang","family":"Xin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wensheng","family":"Yao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"He","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunhao","family":"Hu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rendong","family":"Tong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liangquan","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,6,28]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"616774","DOI":"10.3389\/fvets.2021.616774","article-title":"Transcriptomic analysis of high- and low-virulence bovine pasteurella multocida in vitro and in vivo","volume":"8","author":"He","year":"2021","journal-title":"Front Vet Sci."},{"key":"B2","doi-asserted-by":"publisher","first-page":"280","DOI":"10.1016\/j.vetmic.2019.07.017","article-title":"Antimicrobial resistance of Pasteurella multocida isolated from diseased food-producing animals and pets","volume":"235","author":"Bour\u00e9ly","year":"2019","journal-title":"Vet Microbiol."},{"key":"B3","doi-asserted-by":"publisher","first-page":"301","DOI":"10.3390\/vetsci9060301","article-title":"Spreading of pasteurella multocida infection in a pet rabbit breeding and possible implications on healed bunnies","volume":"9","author":"D'Amico","year":"2022","journal-title":"Vet Sci"},{"key":"B4","author":"Hasan","year":"2022","journal-title":"Pasteurella Multocida"},{"key":"B5","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1089\/vbz.2020.2643","article-title":"Fulminant Septic Shock with Pasteurella multocida in a Young Infant: No Bite, Scratch, or Lick! 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