{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T16:07:15Z","timestamp":1785773235639,"version":"3.56.0"},"reference-count":35,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T00:00:00Z","timestamp":1762473600000},"content-version":"vor","delay-in-days":310,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,1,1]],"date-time":"2025-01-01T00:00:00Z","timestamp":1735689600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2023YFD1800504"],"award-info":[{"award-number":["2023YFD1800504"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012246","name":"Priority Academic Program Development of Jiangsu Higher Education Institutions","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100012246","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Transboundary and Emerging Diseases"],"published-print":{"date-parts":[[2025,1]]},"abstract":"<jats:p>The devastating swine disease, porcine reproductive and respiratory syndrome (PRRS), can only be caused by live PRRS virus (PRRSV) infection. However, the most commonly used detection methods cannot discriminate PRRSV infectivity. Here we developed a triplex propidium monoazide (PMA) qPCR assay for differential detection of infectious PRRSV isolates (NADC34\u2010like PRRSV\u20102, NADC30\u2010like PRRSV\u20102, and HP\u2010PRRSV\u20102) prevalent in China. First, the PRRSV inactivation strategy was selected by comparing distinct inactivation methods. Subsequently, we optimized PMA pretreatment parameters and concentrations of primers and probes. The triplex PMA\u2010qPCR assay displayed favorable specificity, sensitivity, and reproducibility. Moreover, 452 clinical samples (environmental feces, lungs, lymph nodes (LNs), and sera) were submitted to differential detection by triplex qPCR and triplex PMA\u2010qPCR assays. A total of 83 PRRSV\u2010positive samples were detected by the triplex qPCR assay, including 25 NADC34\u2010like, 48 NADC30\u2010like, and 15HP\u2010PRRSV\u20102\u2010positive samples (two samples were coinfected by NADC34\u2010like and NADC30\u2010like PRRSV\u20102, while three samples were coinfected by NADC30\u2010like and HP\u2010PRRSV\u20102). Meanwhile, 65 samples were identified by the PMA\u2010qPCR method, including 21 NADC34\u2010like, 36 NADC30\u2010like, and 9HP\u2010PRRSV\u20102 positive samples (one sample was coinfected by NADC34\u2010like and NADC30\u2010like PRRSV\u20102). No PRRSV could be isolated from the 18 qPCR\u2010positive but PMA\u2010qPCR\u2010negative samples. Overall, this study provides the first triplex PMA\u2010qPCR assay for rapid discrimination of live PRRSV isolates in clinical samples, particularly in environmental feces.<\/jats:p>","DOI":"10.1155\/tbed\/7921675","type":"journal-article","created":{"date-parts":[[2025,11,8]],"date-time":"2025-11-08T06:58:28Z","timestamp":1762585108000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["A Triplex Propidium Monoazide (PMA) qPCR Assay Enables Rapid Discrimination of Live Porcine Reproductive and Respiratory Syndrome Viruses"],"prefix":"10.1155","volume":"2025","author":[{"given":"Xiaoyang","family":"Zhu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenhao","family":"Qi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong","family":"Lin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuan","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuejia","family":"Qiu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ming","family":"Qiu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meng","family":"Cui","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuai","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanhan","family":"Lin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yifan","family":"Meng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wanglong","family":"Zheng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianzhong","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-0030-4217","authenticated-orcid":false,"given":"Zeji","family":"Lu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-3412-5572","authenticated-orcid":false,"given":"Kewei","family":"Fan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1909-8760","authenticated-orcid":false,"given":"Nanhua","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,11,7]]},"reference":[{"key":"e_1_2_11_1_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.ado9328"},{"key":"e_1_2_11_2_2","doi-asserted-by":"publisher","DOI":"10.1177\/104063879200400201"},{"key":"e_1_2_11_3_2","doi-asserted-by":"publisher","DOI":"10.1080\/01652176.1991.9694296"},{"key":"e_1_2_11_4_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-019-04306-w"},{"key":"e_1_2_11_5_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-016-3136-9"},{"key":"e_1_2_11_6_2","first-page":"1","article-title":"Isolation and Identification of Porcine Reproductive and Respiratory Syndrome (PRRS) Virus From Aborted Fetuses Suspected of PRRS","volume":"2","author":"Guo B.","year":"1996","journal-title":"Chinese Journal of Preventive Veterinary Medicine (In Chinese)"},{"key":"e_1_2_11_7_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0000526"},{"key":"e_1_2_11_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01446-15"},{"key":"e_1_2_11_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2018.06.017"},{"key":"e_1_2_11_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2024.110252"},{"key":"e_1_2_11_11_2","doi-asserted-by":"publisher","DOI":"10.1177\/104063870501700211"},{"key":"e_1_2_11_12_2","doi-asserted-by":"publisher","DOI":"10.2174\/1874357901711010066"},{"key":"e_1_2_11_13_2","doi-asserted-by":"publisher","DOI":"10.1111\/tbed.13276"},{"key":"e_1_2_11_14_2","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2025.1616898"},{"key":"e_1_2_11_15_2","doi-asserted-by":"publisher","DOI":"10.1111\/tbed.13368"},{"key":"e_1_2_11_16_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-021-05349-8"},{"key":"e_1_2_11_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.rvsc.2018.10.005"},{"key":"e_1_2_11_18_2","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines9060594"},{"key":"e_1_2_11_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2010.06.011"},{"key":"e_1_2_11_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2015.02.020"},{"key":"e_1_2_11_21_2","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2022.931392"},{"key":"e_1_2_11_22_2","doi-asserted-by":"publisher","DOI":"10.1155\/tbed\/6250851"},{"key":"e_1_2_11_23_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-023-12611-3"},{"key":"e_1_2_11_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2025.110367"},{"key":"e_1_2_11_25_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.00048-25"},{"key":"e_1_2_11_26_2","doi-asserted-by":"publisher","DOI":"10.1186\/1746-6148-6-30"},{"key":"e_1_2_11_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2010.07.016"},{"key":"e_1_2_11_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijbiomac.2025.139670"},{"key":"e_1_2_11_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2024.110213"},{"key":"e_1_2_11_30_2","doi-asserted-by":"publisher","DOI":"10.1002\/fsn3.2916"},{"key":"e_1_2_11_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2006.04.015"},{"key":"e_1_2_11_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.scitotenv.2021.149085"},{"key":"e_1_2_11_33_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2022.1062544"},{"key":"e_1_2_11_34_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaz1439"},{"key":"e_1_2_11_35_2","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2022.882971"}],"container-title":["Transboundary and Emerging Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1155\/tbed\/7921675","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1155\/tbed\/7921675","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1155\/tbed\/7921675","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,8]],"date-time":"2026-03-08T20:59:28Z","timestamp":1773003568000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1155\/tbed\/7921675"}},"subtitle":[],"editor":[{"given":"Linzhu","family":"Ren","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2025,1]]},"references-count":35,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2025,1]]}},"alternative-id":["10.1155\/tbed\/7921675"],"URL":"https:\/\/doi.org\/10.1155\/tbed\/7921675","archive":["Portico"],"relation":{},"ISSN":["1865-1674","1865-1682"],"issn-type":[{"value":"1865-1674","type":"print"},{"value":"1865-1682","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,1]]},"assertion":[{"value":"2025-06-16","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-10-06","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-11-07","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"7921675"}}