{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T23:50:28Z","timestamp":1789429828501,"version":"build-2803163510"},"reference-count":51,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,5,21]],"date-time":"2025-05-21T00:00:00Z","timestamp":1747785600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p>\n                    Sulfonamide resistance genes (\n                    <jats:italic>sul<\/jats:italic>\n                    genes) have a high detection rate and strong transmissibility. Therefore, there is an urgent need to develop more efficient detection methods to enhance the monitoring of\n                    <jats:italic>sul<\/jats:italic>\n                    genes. Current analytical methods are insufficient for the simultaneous and accurate quantification of all sulfonamides resistance genes. To overcome this limitation, a quadruple method was established by integrating droplet digital PCR (ddPCR) with the ratio-based probe-mixing strategy, achieving sensitive detection of\n                    <jats:italic>sul1<\/jats:italic>\n                    ,\n                    <jats:italic>sul2<\/jats:italic>\n                    ,\n                    <jats:italic>sul3<\/jats:italic>\n                    , and\n                    <jats:italic>sul4<\/jats:italic>\n                    genes in diverse matrices. Correspondingly, the primers and probes of\n                    <jats:italic>sul<\/jats:italic>\n                    genes were meticulously designed and rigorously validated, and the critical parameters for ddPCR such as annealing temperature, concentrations of primers and probes were systematically optimized. As a results, the quadruple ddPCR method demonstrates excellent sensitivity with limits of detection (LOD) ranging from 3.98 to 6.16 copies\/reaction, and good repeatability (coefficient of variation &amp;lt;25%), adequately meeting the requirement for accurate\n                    <jats:italic>sul<\/jats:italic>\n                    genes quantification. Furthermore, this new method was applied across 115 diverse samples, including human feces, animal-derived foods, sewage and surface water, achieving positive rates of 100% for\n                    <jats:italic>sul1<\/jats:italic>\n                    , 99.13% for\n                    <jats:italic>sul2<\/jats:italic>\n                    , 93.91% for\n                    <jats:italic>sul3<\/jats:italic>\n                    , and 68.70% for\n                    <jats:italic>sul4<\/jats:italic>\n                    , with\n                    <jats:italic>sul<\/jats:italic>\n                    genes concentration ranging from non-detection to 2.14\u202f\u00d7\u202f10\n                    <jats:sup>9<\/jats:sup>\n                    copies\/g. In summary, the developed quadruple ddPCR method has potential to serve as an efficient and sensitive tool for monitoring\n                    <jats:italic>sul<\/jats:italic>\n                    genes.\n                  <\/jats:p>","DOI":"10.3389\/fmicb.2025.1612740","type":"journal-article","created":{"date-parts":[[2025,5,21]],"date-time":"2025-05-21T01:23:30Z","timestamp":1747790610000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":5,"title":["Development and application of a highly sensitive quadruple droplet digital PCR method for simultaneous quantification of sulfonamide resistance genes"],"prefix":"10.3389","volume":"16","author":[{"given":"Xirong","family":"Yin","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiayuan","family":"Nie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huifang","family":"Tian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lihong","family":"Duan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lixia","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiangdong","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yumei","family":"Guo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ke","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,5,21]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"e0208269","DOI":"10.1371\/journal.pone.0208269","article-title":"High abundances of class 1 integrase and sulfonamide resistance genes, and characterisation of class 1 integron gene cassettes in four urban wetlands in Nigeria","volume":"13","author":"Adelowo","year":"2018","journal-title":"PLoS One"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"886","DOI":"10.1016\/j.scitotenv.2018.12.256","article-title":"Anthropic impacts on sub-Saharan urban water resources through their pharmaceutical contamination (Yaound\u00e9, Center Region, Cameroon)","volume":"660","author":"Branchet","year":"2019","journal-title":"Sci. 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