{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,13]],"date-time":"2026-09-13T19:57:16Z","timestamp":1789329436235,"version":"build-2803163510"},"reference-count":30,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2023,5,15]],"date-time":"2023-05-15T00:00:00Z","timestamp":1684108800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["MicrobiologyOpen"],"published-print":{"date-parts":[[2023,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p><jats:italic>Glaesserella parasuis<\/jats:italic>, <jats:italic>Mycoplasma hyorhinis<\/jats:italic>, and <jats:italic>Mycoplasma hyosynoviae<\/jats:italic> are important porcine pathogens responsible for polyserositis, polyarthritis, meningitis, pneumonia, and septicemia causing significant economic losses in the swine industry. A new multiplex quantitative polymerase chain reaction (qPCR) was designed on one hand for the detection of <jats:italic>G. parasuis<\/jats:italic> and the virulence marker <jats:italic>vtaA<\/jats:italic> to distinguish between highly virulent and non\u2010virulent strains. On the other hand, fluorescent probes were established for the detection and identification of both <jats:italic>M. hyorhinis<\/jats:italic> and <jats:italic>M. hyosynoviae<\/jats:italic> targeting 16S ribosomal RNA genes. The development of the qPCR was based on reference strains of 15 known serovars of <jats:italic>G. parasuis<\/jats:italic>, as well as on the type strains <jats:italic>M. hyorhinis<\/jats:italic> ATCC 17981<jats:sup>T<\/jats:sup> and <jats:italic>M. hyosynoviae<\/jats:italic> NCTC 10167<jats:sup>T<\/jats:sup>. The new qPCR was further evaluated using 21 <jats:italic>G. parasuis<\/jats:italic>, 26 <jats:italic>M. hyorhinis<\/jats:italic>, and 3\u00a0<jats:italic>M. hyosynoviae<\/jats:italic> field isolates. Moreover, a pilot study including different clinical specimens of 42 diseased pigs was performed. The specificity of the assay was 100% without cross\u2010reactivity or detection of other bacterial swine pathogens. The sensitivity of the new qPCR was demonstrated to be between 11\u2013180 genome equivalents (GE) of DNA for <jats:italic>M. hyosynoviae<\/jats:italic> and <jats:italic>M. hyorhinis<\/jats:italic>, and 140\u20131200 GE for <jats:italic>G. parasuis<\/jats:italic> and <jats:italic>vtaA<\/jats:italic>. The cut\u2010off threshold cycle was found to be at 35. The developed sensitive and specific qPCR assay has the potential to become a useful molecular tool, which could be implemented in veterinary diagnostic laboratories for the detection and identification of <jats:italic>G. parasuis<\/jats:italic>, its virulence marker <jats:italic>vtaA<\/jats:italic>, <jats:italic>M. hyorhinis<\/jats:italic>, and <jats:italic>M. hyosynoviae<\/jats:italic>.<\/jats:p>","DOI":"10.1002\/mbo3.1353","type":"journal-article","created":{"date-parts":[[2023,5,16]],"date-time":"2023-05-16T06:09:34Z","timestamp":1684217374000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Development of a new multiplex quantitative PCR for the detection of <i>Glaesserella parasuis<\/i>, <i>Mycoplasma hyorhinis<\/i>, and <i>Mycoplasma hyosynoviae<\/i>"],"prefix":"10.1002","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9548-8798","authenticated-orcid":false,"given":"Simone","family":"Scherrer","sequence":"first","affiliation":[{"name":"Section of Veterinary Bacteriology, Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich Zurich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah","family":"Schmitt","sequence":"additional","affiliation":[{"name":"Section of Veterinary Bacteriology, Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich Zurich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fenja","family":"Rademacher","sequence":"additional","affiliation":[{"name":"Section of Veterinary Bacteriology, Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich Zurich 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