{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T09:57:13Z","timestamp":1789811833097,"version":"4.0.1"},"reference-count":34,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2025,2,11]],"date-time":"2025-02-11T00:00:00Z","timestamp":1739232000000},"content-version":"vor","delay-in-days":41,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82270593"],"award-info":[{"award-number":["82270593"]}],"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":["82200628"],"award-info":[{"award-number":["82200628"]}],"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":["82060109"],"award-info":[{"award-number":["82060109"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Helicobacter"],"published-print":{"date-parts":[[2025,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:sec><jats:title>Aims<\/jats:title><jats:p>This study evaluates the accuracy of melting curve\u2013based multiplex real\u2010time PCR (multiplex rt\u2010PCR) on stool samples for diagnosing antibiotic resistance in <jats:styled-content style=\"fixed-case\"><jats:italic>Helicobacter pylori<\/jats:italic><\/jats:styled-content> (<jats:styled-content style=\"fixed-case\"><jats:italic>H. pylori<\/jats:italic><\/jats:styled-content>) compared to E\u2010test and sequencing.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Gastric biopsies and stool samples were collected from 385 <jats:styled-content style=\"fixed-case\"><jats:italic>H. pylori<\/jats:italic><\/jats:styled-content>\u2010infected patients. A total of 325 strains were isolated, and genomic DNA was extracted from all 385 stool samples. E\u2010tests were conducted to detect phenotypic resistance for clarithromycin and levofloxacin. Sanger sequencing and multiplex rt\u2010PCR were employed to identify <jats:styled-content style=\"fixed-case\"><jats:italic>H. pylori<\/jats:italic><\/jats:styled-content> 23S rRNA and GyrA mutations.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>E\u2010test results indicated that 203 (62.5%) were susceptible to both clarithromycin and levofloxacin, 33 (10.2%) exhibited mono\u2010resistance to clarithromycin, 48 (14.8%) showed mono\u2010resistance to levofloxacin, and 41 (12.6%) had dual resistance to both antibiotics. Compared to E\u2010test results, the sensitivity and specificity of the multiplex rt\u2010PCR method for detecting clarithromycin resistance mutation were 93.2 (95% CI 84.3\u201397.5) and 87.1% (95% CI 82.2\u201390.9), respectively. For levofloxacin resistance mutation, the multiplex rt\u2010PCR method showed a sensitivity of 80.7 (95% CI 70.3\u201388.3) and a specificity of 93.0% (95% CI 88.7\u201395.8). Compared to Sanger sequencing, the sensitivity and specificity of the multiplex rt\u2010PCR method for detecting clarithromycin resistance mutation were 95.8 (95% CI 90.0\u201398.4) and 96.0% (95% CI 92.6\u201398.0), respectively. For levofloxacin resistance mutation, the multiplex rt\u2010PCR method showed a sensitivity of 91.3% (95% CI, 83.1\u201395.9) and a specificity of 96.1% (95% CI, 92.7\u201398.0).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Genotypic methods, including Sanger sequencing and multiplex rt\u2010PCR, were rapid and reliable for diagnosing clarithromycin and levofloxacin resistance in the stool samples.<\/jats:p><\/jats:sec>","DOI":"10.1111\/hel.70012","type":"journal-article","created":{"date-parts":[[2025,2,12]],"date-time":"2025-02-12T03:45:14Z","timestamp":1739331914000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["The Accuracy of Melting Curve\u2013Based Multiplex Real\u2010Time <scp>PCR<\/scp> for Diagnosing <scp><i>Helicobacter pylori<\/i><\/scp> Resistance to Clarithromycin and Levofloxacin in Stool Specimens"],"prefix":"10.1111","volume":"30","author":[{"given":"Youhua","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Gastroenterology, Jiangxi Provincial Key Laboratory of Digestive Diseases, Jiangxi Clinical Research Center for Gastroenterology, Digestive Disease Hospital, the First Affiliated Hospital, Jiangxi Medical College Nanchang University  Nanchang Jiangxi China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoling","family":"Gong","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Jiangxi Provincial Key Laboratory of Digestive Diseases, Jiangxi Clinical Research Center for Gastroenterology, Digestive Disease Hospital, the First Affiliated Hospital, Jiangxi Medical College Nanchang University  Nanchang Jiangxi China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qirui","family":"Lai","sequence":"additional","affiliation":[{"name":"Huan Kui College of Nanchang University Jiangxi Medical College, Nanchang University  Nanchang Jiangxi Province China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Fang","sequence":"additional","affiliation":[{"name":"Huan Kui College of Nanchang University Jiangxi Medical College, Nanchang University  Nanchang Jiangxi Province China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiyin","family":"Yang","sequence":"additional","affiliation":[{"name":"Jiangsu GeneWill Diagnostics Company Limited  Jiangsu Province China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuan","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Jiangxi Provincial Key Laboratory of Digestive Diseases, Jiangxi Clinical Research Center for Gastroenterology, Digestive Disease Hospital, the First Affiliated Hospital, Jiangxi Medical College Nanchang University  Nanchang Jiangxi China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dong Sheng","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Jiangxi Provincial Key Laboratory of Digestive Diseases, Jiangxi Clinical Research Center for Gastroenterology, Digestive Disease Hospital, the First Affiliated Hospital, Jiangxi Medical College Nanchang University  Nanchang Jiangxi China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5290-5579","authenticated-orcid":false,"given":"Yong","family":"Xie","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Jiangxi Provincial Key Laboratory of Digestive Diseases, Jiangxi Clinical Research Center for Gastroenterology, Digestive Disease Hospital, the First Affiliated Hospital, Jiangxi Medical College Nanchang University  Nanchang Jiangxi 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