{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,24]],"date-time":"2026-07-24T02:40:00Z","timestamp":1784860800259,"version":"3.55.0"},"reference-count":56,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2024,4,19]],"date-time":"2024-04-19T00:00:00Z","timestamp":1713484800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Guangxi Key Laboratory of Animal Breeding, Disease Control, and Prevention","award":["ABDC-b202304"],"award-info":[{"award-number":["ABDC-b202304"]}]},{"name":"Guangxi Key Laboratory of Animal Breeding, Disease Control, and Prevention","award":["AB21238003"],"award-info":[{"award-number":["AB21238003"]}]},{"name":"Key Research and Development Program of Guangxi Science and Technology Bureau, China","award":["ABDC-b202304"],"award-info":[{"award-number":["ABDC-b202304"]}]},{"name":"Key Research and Development Program of Guangxi Science and Technology Bureau, China","award":["AB21238003"],"award-info":[{"award-number":["AB21238003"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Porcine respiratory coronavirus (PRCoV), porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), and pseudorabies virus (PRV) are significant viruses causing respiratory diseases in pigs. Sick pigs exhibit similar clinical symptoms such as fever, cough, runny nose, and dyspnea, making it very difficult to accurately differentially diagnose these diseases on site. In this study, a quadruplex one-step reverse-transcription real-time quantitative PCR (RT-qPCR) for the detection of PRCoV, PRRSV, SIV, and PRV was established. The assay showed strong specificity, high sensitivity, and good repeatability. It could detect only PRCoV, PRRSV, SIV, and PRV, without cross-reactions with TGEV, PEDV, PRoV, ASFV, FMDV, PCV2, PDCoV, and CSFV. The limits of detection (LODs) for PRCoV, PRRSV, SIV, and PRV were 129.594, 133.205, 139.791, and 136.600 copies\/reaction, respectively. The intra-assay and inter-assay coefficients of variation (CVs) ranged from 0.29% to 1.89%. The established quadruplex RT-qPCR was used to test 4909 clinical specimens, which were collected in Guangxi Province, China, from July 2022 to September 2023. PRCoV, PRRSV, SIV, and PRV showed positivity rates of 1.36%, 10.17%, 4.87%, and 0.84%, respectively. In addition, the previously reported RT-qPCR was also used to test these specimens, and the agreement between these methods was higher than 99.43%. The established quadruplex RT-qPCR can accurately detect these four porcine respiratory viruses simultaneously, providing an accurate and reliable detection technique for clinical diagnosis.<\/jats:p>","DOI":"10.3390\/pathogens13040341","type":"journal-article","created":{"date-parts":[[2024,4,19]],"date-time":"2024-04-19T06:28:09Z","timestamp":1713508089000},"page":"341","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Simultaneous Detection of Porcine Respiratory Coronavirus, Porcine Reproductive and Respiratory Syndrome Virus, Swine Influenza Virus, and Pseudorabies Virus via Quadruplex One-Step RT-qPCR"],"prefix":"10.3390","volume":"13","author":[{"given":"Yan","family":"Ma","sequence":"first","affiliation":[{"name":"College of Animal Science and Technology, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1023-9635","authenticated-orcid":false,"given":"Kaichuang","family":"Shi","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Guangxi University, Nanning 530004, China"},{"name":"Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenhai","family":"Chen","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuwen","family":"Shi","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingan","family":"Zhou","sequence":"additional","affiliation":[{"name":"Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shenglan","family":"Mo","sequence":"additional","affiliation":[{"name":"Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haina","family":"Wei","sequence":"additional","affiliation":[{"name":"Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liping","family":"Hu","sequence":"additional","affiliation":[{"name":"Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3903-0509","authenticated-orcid":false,"given":"Meilan","family":"Mo","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,4,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e01355-20","DOI":"10.1128\/JCM.01355-20","article-title":"Comparative pathogenesis of bovine and porcine respiratory coronaviruses in the animal host species and SARS-CoV-2 in humans","volume":"58","author":"Saif","year":"2020","journal-title":"J. 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