{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T00:53:18Z","timestamp":1760057598927,"version":"build-2065373602"},"reference-count":43,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2025,2,14]],"date-time":"2025-02-14T00:00:00Z","timestamp":1739491200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Capital\u2019s Funds for Health Improvement and Research","award":["CFH 2024-1G-4362"],"award-info":[{"award-number":["CFH 2024-1G-4362"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The abuse of antibiotics has led to widespread resistance to Helicobacter pylori (H. pylori) in the population. There is an urgent need to establish a method to detect multiple antibiotic resistance rapidly. This study aimed to construct a novel strategy for the high-throughput detection of H. pylori\u2019s resistance to varying antibiotics using multiplex PCR\u2013mass spectrometry mini-sequencing (mPCR-MS mini-sequencing) technology. This study detected the resistance of H. pylori to six antibiotics using eight mutated sites (23S rRNA-2143; pbp1A-1667, 1684, 1240; gyrA-261, 271, 573; and 16S rRNA-928) of four resistance genes (pbp1A, gyrA, 23S rRNA, and 16S rRNA), and 525 were detected in all 528 results (99.43%). Then, the culture-based phenotypic drug susceptibility testing (DST) method was used as a reference for drug resistance detection. We found that the consistency rate between mPCR-MS mini-sequencing with the DST results of amoxicillin (AMX), moxifloxacin (MOX), levofloxacin (LEV), clarithromycin (CLA), azithromycin (AZI), and tetracycline (TET) were 95.5% (63\/66), 77.3% (51\/66), 68.2% (45\/66), 93.9% (62\/66), 92.4% (61\/66), and 97.0% (64\/66), respectively. This method was high-throughput and extensible, easily improving the entire detection system by adding new mutation sites. mPCR-MS mini-sequencing technology provides a new approach to mutation sites related to H. pylori\u2019s antibiotic resistance.<\/jats:p>","DOI":"10.3390\/ijms26041632","type":"journal-article","created":{"date-parts":[[2025,2,14]],"date-time":"2025-02-14T04:48:51Z","timestamp":1739508531000},"page":"1632","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Multiplex PCR\u2013Mass Spectrometry Mini-Sequencing Technology Detected Antibiotic Resistance of Helicobacter pylori to Six Antibiotics"],"prefix":"10.3390","volume":"26","author":[{"given":"Fei","family":"Zhao","sequence":"first","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-4381-4712","authenticated-orcid":false,"given":"Xin","family":"Zhao","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huifang","family":"Zhang","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lihua","family":"He","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fanliang","family":"Meng","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7056-8206","authenticated-orcid":false,"given":"Jianzhong","family":"Zhang","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5964-5925","authenticated-orcid":false,"given":"Di","family":"Xiao","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1053\/j.gastro.2017.04.022","article-title":"Global Prevalence of Helicobacter pylori Infection: Systematic Review and Meta-Analysis","volume":"153","author":"Hooi","year":"2017","journal-title":"Gastroenterology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1038\/s41575-021-00449-x","article-title":"Helicobacter pylori infection and antibiotic resistance\u2014From biology to clinical implications","volume":"18","author":"Yamaoka","year":"2021","journal-title":"Nat. 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