{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T10:45:39Z","timestamp":1782297939022,"version":"3.54.5"},"reference-count":25,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,6,23]],"date-time":"2025-06-23T00:00:00Z","timestamp":1750636800000},"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. Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p><jats:italic>Helicobacter pylori<\/jats:italic> infection poses a significant global health challenge, exacerbated by rising antibiotic resistance. This study aimed to evaluate a novel multiplex RT-PCR-based fecal diagnostic kit (Cowin Biosciences, Jiangsu, China) for detecting mutations in the <jats:italic>23S rRNA<\/jats:italic> and <jats:italic>gyrA<\/jats:italic> genes associated with clarithromycin and fluoroquinolone resistance in <jats:italic>H. pylori<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>A total of 1,176 participants from four clinical centers in China were enrolled between August 2022 and October 2023. Phenotypic resistance was assessed on <jats:italic>H. pylori<\/jats:italic> isolated from gastric samples using the minimum inhibitory concentration (MIC) method (E-test), while fecal samples were analyzed molecularly via the diagnostic kit and Sanger sequencing. Positive (PPA), negative (NPA), and overall percentage agreement (OPA) with 95% confidence intervals (CI) were calculated.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>There was a high level of consistency between phenotypic testing and novel diagnostic kit. The PPA, NPA, and OPA of the fecal diagnostic kit for detecting clarithromycin susceptibility were 92.97% (CI: 90.6%-95.3%), 87.89% (CI: 85.0%-90.8%), and 90.36% (CI: 88.5%-92.3%), respectively. And likewise, the PPA, NPA, and OPA in diagnosing <jats:italic>H<\/jats:italic>. <jats:italic>pylori<\/jats:italic> resistance to fluoroquinolones were 86.85% (CI:83.6%- 90.1%), 91.12% (CI: 88.6%-93.7%), and 89.12% (CI: 87.1%-91.1%), respectively. Beyond that, the diagnostic kit also exhibited a high degree of concordance with the outcomes of Sanger sequencing. Specifically, when assessing clarithromycin resistance, the PPA, NPA, and OPA of the diagnostic kit were 97.13% (CI: 95.7%-98.6%), 94.62% (CI: 92.6%-96.6%), and 95.90% (CI: 94.7%-97.1%), respectively. Similarly, application of the diagnostic kit to detect fluoroquinolone resistance achieved a PPA of 96.92% (CI: 94.9%-98.4%), an NPA of 93.18% (CI: 91.0%-95.3%), and an OPA of 94.69% (CI: 93.3%-96.1%). The concordance rate between the fecal kit and phenotypic susceptibility testing varied with MIC values. For clarithromycin resistance, the positive percentage agreement (PPA) was lowest (89.74%) when MIC exceeded 256 \u00b5g\/mL but peaked at 96.47% for MIC values between 32 and 256 \u00b5g\/mL. The highest overall concordance (Kappa = 0.774) was observed at intermediate MIC levels (4\u201332 \u00b5g\/mL), suggesting optimal detection accuracy in this range.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The fecal diagnostic kit provides a rapid, non-invasive, and reliable method to predict clarithromycin and fluoroquinolone resistance, supporting personalized therapy.<\/jats:p><\/jats:sec><jats:sec><jats:title>Clinical Trial Registration<\/jats:title><jats:p>ClinicalTrials.gov, identifier NCT05410652.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2025.1592612","type":"journal-article","created":{"date-parts":[[2025,6,23]],"date-time":"2025-06-23T05:21:18Z","timestamp":1750656078000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Clinical evaluation of a novel H. pylori fecal molecular diagnosis kit (multiplex RT-PCR method) for detecting clarithromycin and fluoroquinolones resistance using stool samples"],"prefix":"10.3389","volume":"15","author":[{"given":"Wen-juan","family":"Wei","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bang sun","family":"He","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Lv","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong","family":"Xie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhen yu","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,6,23]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"2400","DOI":"10.1128\/JCM.00506-17","article-title":"A Novel Stool PCR Test for Helicobacter pylori May Predict Clarithromycin Resistance and Eradication of Infection at a High Rate","volume":"55","author":"Beckman","year":"2017","journal-title":"J. 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