{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,19]],"date-time":"2026-01-19T12:40:42Z","timestamp":1768826442116,"version":"3.49.0"},"reference-count":41,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,9,6]],"date-time":"2022-09-06T00:00:00Z","timestamp":1662422400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32102669"],"award-info":[{"award-number":["32102669"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31972685,"],"award-info":[{"award-number":["31972685,"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31920103015"],"award-info":[{"award-number":["31920103015"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004608","name":"Natural Science Foundation of Jiangsu Province","doi-asserted-by":"publisher","award":["BK20210802"],"award-info":[{"award-number":["BK20210802"]}],"id":[{"id":"10.13039\/501100004608","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002858","name":"China Postdoctoral Science Foundation","doi-asserted-by":"publisher","award":["2020M681748"],"award-info":[{"award-number":["2020M681748"]}],"id":[{"id":"10.13039\/501100002858","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010246","name":"Postdoctoral Science Foundation of Jiangsu Province","doi-asserted-by":"publisher","award":["2021K095A"],"award-info":[{"award-number":["2021K095A"]}],"id":[{"id":"10.13039\/501100010246","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p><jats:italic>Salmonella enterica<\/jats:italic> serovar Gallinarum biovars Gallinarum and Pullorum cause severe chicken salmonellosis, a disease associated with high mortality and morbidity among chickens worldwide. The conventional serotyping and biochemical reactions have been used to identify <jats:italic>Salmonella<\/jats:italic> serovars. However, the conventional methods are complicated, time-consuming, laborious, and expensive. Furthermore, it is challenging to distinguish <jats:italic>S.<\/jats:italic> Gallinarum and <jats:italic>S.<\/jats:italic> Pullorum <jats:italic>via<\/jats:italic> biochemical assays and serotyping because of their antigenic similarity. Although various PCR methods were established, a PCR protocol to detect and discriminate <jats:italic>S.<\/jats:italic> Gallinarum and <jats:italic>S.<\/jats:italic> Pullorum simultaneously is lacking. Herein, a one-step multiplex PCR method was established for the accurate identification and discrimination of <jats:italic>S.<\/jats:italic> Pullorum and <jats:italic>S.<\/jats:italic> Gallinarum. Three specific genes were used for the multiplex PCR method, with the <jats:italic>I137_14445<\/jats:italic> and <jats:italic>ybgL<\/jats:italic> genes being the key targets to identify and differentiate <jats:italic>S.<\/jats:italic> Gallinarum <jats:italic>and S.<\/jats:italic> Pullorum, and <jats:italic>stn<\/jats:italic> being included as a reference gene for the <jats:italic>Salmonella<\/jats:italic> genus. <jats:italic>In silico<\/jats:italic> analysis showed that the <jats:italic>I137_14445<\/jats:italic> gene is present in all <jats:italic>Salmonella<\/jats:italic> serovars, except for <jats:italic>S.<\/jats:italic> Gallinarum, and could therefore be used for the identification of <jats:italic>S.<\/jats:italic> Gallinarum. A 68-bp sequence deficiency in <jats:italic>ybgL<\/jats:italic> was found only in <jats:italic>S.<\/jats:italic> Pullorum compared to other <jats:italic>Salmonella<\/jats:italic> serovars, and this could therefore be used for the specific identification of <jats:italic>S.<\/jats:italic> Pullorum. The developed PCR assay was able to distinguish <jats:italic>S.<\/jats:italic> Gallinarum <jats:italic>and S.<\/jats:italic> Pullorum among 75 various <jats:italic>Salmonella<\/jats:italic> strains and 43 various non-<jats:italic>Salmonella<\/jats:italic> pathogens with excellent specificity. The detection limit for the genomic DNA of <jats:italic>S.<\/jats:italic> Gallinarum and <jats:italic>S.<\/jats:italic> Pullorum was 21.4\u2009pg.\/\u03bcL, and the detectable limit for bacterial cells was 100\u2009CFU. The developed PCR method was used for the analysis of <jats:italic>Salmonella<\/jats:italic> isolates in a chicken farm. This PCR system successfully discriminated <jats:italic>S.<\/jats:italic> Gallinarum <jats:italic>and S.<\/jats:italic> Pullorum from other different <jats:italic>Salmonella<\/jats:italic> serovars. The PCR results were confirmed by the conventional serotyping method. The newly established multiplex PCR is a simple, accurate, and cost-effective method for the timely identification and differentiation of <jats:italic>S.<\/jats:italic> Pullorum and <jats:italic>S.<\/jats:italic> Gallinarum.<\/jats:p>","DOI":"10.3389\/fmicb.2022.983942","type":"journal-article","created":{"date-parts":[[2022,9,6]],"date-time":"2022-09-06T05:26:56Z","timestamp":1662442016000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["A new multiplex PCR for the accurate identification and differentiation of Salmonella enterica serovar Gallinarum biovars Pullorum and Gallinarum"],"prefix":"10.3389","volume":"13","author":[{"given":"Dan","family":"Xiong","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Li","family":"Yuan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Li","family":"Song","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xinan","family":"Jiao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhiming","family":"Pan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2022,9,6]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"109516","DOI":"10.1016\/j.ijfoodmicro.2021.109516","article-title":"Evidence of sheep and abattoir environment as important reservoirs of multidrug resistant salmonella and extended-spectrum beta-lactamase Escherichia coli","volume":"363","author":"Atlaw","year":"2022","journal-title":"Int. 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