{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T18:01:09Z","timestamp":1785261669015,"version":"3.55.0"},"reference-count":25,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T00:00:00Z","timestamp":1785196800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"General Research Project of Zhejiang Provincial Education Department","award":["Y202559640"],"award-info":[{"award-number":["Y202559640"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    <jats:italic>Mycoplasma synoviae<\/jats:italic>\n                    (MS) is a significant avian pathogen responsible for arthritis, tenosynovitis, airsacculitis, and abnormal eggshell apex syndrome in chickens, posing a substantial threat to the poultry industry. While the attenuated MS-H vaccine has proven effective and is widely implemented in poultry flocks. However, distinguishing the MS-H vaccine strain from wild-type strains remains a persistent challenge. Recently, Pyrococcus furiosus Argonaute (PfAgo) nucleases have garnered considerable attention due to their capacity for single-nucleotide discrimination. Leveraging the A367G SNP within the MS\n                    <jats:italic>obg<\/jats:italic>\n                    gene, we developed a novel identification method that integrates PfAgo-mediated cleavage with PCR amplification. Through systematic optimization of PfAgo cleavage substrates and PCR primers, this approach achieved detection sensitivities of 1\u2009\u00d7\u200910\n                    <jats:sup>3<\/jats:sup>\n                    copies\/\u00b5L for the MS-H vaccine strain and 1\u2009\u00d7\u200910\n                    <jats:sup>4<\/jats:sup>\n                    copies\/\u00b5L for wild-type strains. Validation using 12 clinical samples resulted in the accurate identification of three MS-H vaccine strains and nine wild-type strains. The established method thus provides a reliable and sensitive tool for discriminating between MS-H vaccine and wild-type strains, supporting improved surveillance and control in poultry farming.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0351464","type":"journal-article","created":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T17:28:04Z","timestamp":1785259684000},"page":"e0351464","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Pyrococcus furiosus Argonaute coupled PCR assay for accurate discrimination between the MS-H vaccine strain and clinical isolates of Mycoplasma synoviae"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1243-1436","authenticated-orcid":true,"given":"Yanli","family":"Zhao","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yinling","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ge","family":"Song","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Houqiang","family":"Luo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liyan","family":"Dong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingsong","family":"Han","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengling","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Pan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8349-2877","authenticated-orcid":true,"given":"Hongxia","family":"Jiang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,7,28]]},"reference":[{"issue":"3","key":"pone.0351464.ref001","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1637\/8323-041808-Review.1","article-title":"Control 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