{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,19]],"date-time":"2026-10-19T23:01:48Z","timestamp":1792450908284,"version":"4.5.0"},"reference-count":2,"publisher":"Polish Academy of Sciences Chancellery","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:p>The control of respiratory infections is integral to a healthy livestock farm. The current study was conducted to optimize a multiplex PCR assay for simultaneous detection of Pasteurella multocida (P. multocida), Staphylococcus aureus (S. aureus) and Mycobacterium bovis (M. bovis) in nasal samples of cattle having suspicion of respiratory tract infections. The nasal samples were collected from 10 dairy farms affected with respiratory disease outbreaks in the recent past. These outbreaks were associated with respiratory tract infections caused by bacterial pathogens P. multocida, S. aureus and M. bovis. A multiplex PCR assay was therefore optimized to enable the simultaneous detection of these bacterial pathogens directly from nasal samples. The multi\u00adplex PCR assay was optimized using primers already validated for efficient amplification of spe\u00adcific DNA segments in reference strains of targeted bacterial pathogens. The standardized assay was specific and sensitive enough to detect \u2265100 genome equivalents of target DNA segments in each of P. multocida, S. aureus and M. bovis. The assay was successfully applied for the detec\u00adtion of the three bacterial pathogens in 50 nasal samples. PCR amplicons were subjected to Sanger dideoxy sequencing followed by phylogenetic analysis to determine if species identifi\u00adcation was specific. In short, the optimized multiplex PCR assay may prove as a reliable, econo-mical, and simple tool for the management of bovine respiratory tract infections caused by some key bacterial pathogens.<\/jats:p>","DOI":"10.24425\/pjvs.2025.154015","type":"journal-article","created":{"date-parts":[[2025,3,20]],"date-time":"2025-03-20T11:13:30Z","timestamp":1742469210000},"page":"73-73","source":"Crossref","is-referenced-by-count":3,"title":["Optimization of a multiplex PCR assay for simultaneous detection of key bovine respiratory infections"],"prefix":"10.24425","author":[{"given":"M.","family":"Jamil","sequence":"first","affiliation":[{"name":"Dept. of Wildlife and Ecology, University of Veterinary and Animal Sciences, 54600-Lahore-Pakistan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S.","family":"Sidra","sequence":"additional","affiliation":[{"name":"Dept. of Wildlife and Ecology, University of Veterinary and Animal Sciences, 54600-Lahore-Pakistan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.","family":"Hussain","sequence":"additional","affiliation":[{"name":"Institute of Zoology, University of the Punjab, 54600-Lahore-Pakistan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Imran","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, 54600-Lahore-Pakistan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.A.","family":"Sheikh","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, University of Veterinary and Animal Sciences, 54600-Lahore-Pakistan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"37468","published-online":{"date-parts":[[2025,3,20]]},"reference":[{"key":"#cr-split#-ref0.1","unstructured":"Ashraf A, Imran M, Yaqub T, Tayyab M, Shehzad W,Thomson PC (2017) A novel multiplex PCR assay forsimultaneous detection of nine clinically significant bacterial pathogens associated with bovine mastitis. 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Vet Clin N Am Food A 31: 351-365."}],"container-title":["Polish Journal of Veterinary Sciences"],"original-title":[],"language":"pl","link":[{"URL":"https:\/\/journals.pan.pl\/Content\/134455\/PDF\/8%20_%20Jamil.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,3,20]],"date-time":"2025-03-20T14:21:27Z","timestamp":1742480487000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.pan.pl\/dlibra\/publication\/154015\/edition\/134455\/content"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,20]]},"references-count":2,"URL":"https:\/\/doi.org\/10.24425\/pjvs.2025.154015","relation":{},"ISSN":["1505-1773"],"issn-type":[{"value":"1505-1773","type":"print"}],"subject":[],"published":{"date-parts":[[2025,3,20]]}}}