{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T11:22:29Z","timestamp":1785410549191,"version":"3.56.0"},"reference-count":36,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2026,5,20]],"date-time":"2026-05-20T00:00:00Z","timestamp":1779235200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,5,20]],"date-time":"2026-05-20T00:00:00Z","timestamp":1779235200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100005825","name":"National Institute of Food and Agriculture","doi-asserted-by":"publisher","award":["2022-6801536628"],"award-info":[{"award-number":["2022-6801536628"]}],"id":[{"id":"10.13039\/100005825","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2026,5,20]]},"abstract":"ABSTRACT<\/jats:title>\n \n \n \n Salmonella<\/jats:italic>\n enterica serovar Dublin (\n S<\/jats:italic>\n . Dublin) is a host-adapted cattle pathogen that causes systemic disease, chronic carriage, and environmental contamination. Early detection of subclinical infections remains challenging due to low bacterial loads and the limitations of conventional diagnostics. We developed a duplex TaqMan real-time PCR assay targeting a chromosomal locus (SeD_A1104) and a plasmid-encoded gene (\n vagD<\/jats:italic>\n ). The assay demonstrated strong linearity (\n R<\/jats:italic>\n \u00b2 > 0.99) with amplification efficiencies of 91%\u2013102% and detection limits as low as 1.9 \u00d7 10\u00b9 genome copies per reaction. Specificity was confirmed\n in silico<\/jats:italic>\n and against 20 non-Dublin\n Salmonella<\/jats:italic>\n serovars, with no cross-amplification. Intra- and inter-assay variability was consistently below 3%. Field testing of 40 samples from subclinical cattle and farm environments demonstrated detection in multiple matrices, including feces (50% positive), vaginal swabs (40%), nasal swabs (30%), and environmental boot-swabs (75%). The dual-target design provides improved confidence in detection, reduces false positives, and supports early detection and control of\n S<\/jats:italic>\n . Dublin in cattle herds and associated environments, thereby contributing to reduced zoonotic transmission and enhancing food safety within a One Health framework.\n <\/jats:p>\n \n IMPORTANCE<\/jats:title>\n \n Early detection of\n Salmonella<\/jats:italic>\n Dublin is critical for controlling disease in cattle and reducing zoonotic risk. Existing diagnostics often fail to identify subclinical carriers or low-level environmental contamination, which sustain transmission within herds. We developed a dual-target duplex quantitative real-time PCR assay that simultaneously amplifies a chromosomal locus and a plasmid-encoded gene, improving specificity and reducing false positives caused by genetically related\n Salmonella<\/jats:italic>\n serovars. Unlike many current tools, this assay is validated for direct use with challenging samples, including feces, nasal and vaginal swabs, and environmental boot swabs, where inhibitors and low pathogen load commonly interfere with detection. By enhancing sensitivity and reliability across diverse matrices, this method supports farm-level surveillance, rapid outbreak response, and risk-based control strategies. Broader implementation can improve herd health, limit economic losses, and reduce the risk of human exposure through food or environmental sources.\n <\/jats:p>\n <\/jats:sec>\n <\/jats:sec>","DOI":"10.1128\/aem.01979-25","type":"journal-article","created":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T13:02:33Z","timestamp":1776344553000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Novel dual-gene target and probe-based real-time PCR for the detection and differentiation of\n Salmonella<\/i>\n Dublin"],"prefix":"10.1128","volume":"92","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6679-3785","authenticated-orcid":true,"given":"Alejandra","family":"Arevalo-Mayorga","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00rs6vg23","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Veterinary Preventive Medicine, The Ohio State University","place":["Columbus, 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