{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:19:10Z","timestamp":1785403150085,"version":"3.56.0"},"reference-count":64,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,5,22]],"date-time":"2026-05-22T00:00:00Z","timestamp":1779408000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"crossref"}]},{"id":[{"id":"https:\/\/ror.org\/004rtk039","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Adopting Sustainable Partnerships for Innovative Research Ecosystem","award":["23836904"],"award-info":[{"award-number":["23836904"]}]},{"DOI":"10.13039\/501100009037","name":"Science and Technology Research Partnership for Sustainable Development","doi-asserted-by":"publisher","award":["21357105"],"award-info":[{"award-number":["21357105"]}],"id":[{"id":"10.13039\/501100009037","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001700","name":"Ministry of Education, Culture, Sports, Science and Technology","doi-asserted-by":"publisher","award":["23K24133"],"award-info":[{"award-number":["23K24133"]}],"id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"publisher"}]},{"award":["23K24133"],"award-info":[{"award-number":["23K24133"]}],"id":[{"id":"https:\/\/ror.org\/048rj2z13","id-type":"ROR","asserted-by":"publisher"}]},{"name":"JSPS Core-to-Core Program","award":["G925F01003"],"award-info":[{"award-number":["G925F01003"]}]},{"name":"Research Center for Global and Local Infectious Diseases","award":["2024B26"],"award-info":[{"award-number":["2024B26"]}]},{"name":"Research Center for Global and Local Infectious Diseases","award":["2025B21"],"award-info":[{"award-number":["2025B21"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Accurate detection and monitoring of antimicrobial resistance (AMR) in Helicobacter pylori mainly rely on phenotypic methods and culture, which can sometimes fail when bacterial load is low or after recent treatment. We investigated whether gastric biopsies classified as H. pylori-negative by standard diagnostic techniques still contain detectable bacterial DNA, including regions linked to AMR, and assessed whether selected DNA fragments can mediate allelic exchange in vitro. Gastric biopsies from 46 dyspeptic patients in the Democratic Republic of the Congo (including 23 phenotypically positive and 23 phenotypically negative individuals) were analyzed using long-read amplicon sequencing of seven resistance-associated loci, selective whole-genome amplification (sWGA) followed by long-read sequencing of H. pylori-enriched reads, and a proof-of-concept natural transformation assay. Phenotypically negative biopsies exhibited significantly lower sequencing depth across multiple loci (including 23S rRNA, gyrA, gyrB, and pbp1A; p = 0.003\u20130.014), indicating a reduced H. pylori DNA burden. However, AMR-associated mutations linked to various antibiotic classes were found in both groups. sWGA enabled recovery of fragmentary H. pylori sequence data from phenotypically negative samples, including reads that map to resistance- and virulence-associated genes. In vitro, 23S rRNA A2143G amplicons from both phenotypically positive and negative biopsies produced clarithromycin-resistant transformants in strain 26695. These findings indicate that phenotypically negative gastric biopsies might contain low-abundance and fragmentary H. pylori DNA. Although certain DNA fragments can mediate allelic exchange under controlled in vitro conditions, these results do not confirm bacterial viability, active infection, or clinically relevant in vivo resistance transfer. Therefore, they should be interpreted with caution in molecular AMR surveillance and detection contexts.<\/jats:p>","DOI":"10.3390\/biom16060765","type":"journal-article","created":{"date-parts":[[2026,5,22]],"date-time":"2026-05-22T16:06:08Z","timestamp":1779465968000},"page":"765","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Low-Abundance and Fragmentary Helicobacter pylori DNA Detected in Phenotypically Negative Gastric Biopsies Using Targeted Sequencing"],"prefix":"10.3390","volume":"16","author":[{"given":"Fabien","family":"Mbaya-Tshibangu","sequence":"first","affiliation":[{"name":"Department of Environmental and Preventive Medicine, Faculty of Medicine, Oita University, Yufu 879-5593, Japan"},{"name":"Department of Internal Medicine, Faculty of Medicine, Pharmacy and Public Health, University of Mbujimayi, Mbujimayi P.O. Box 225, Congo"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alain","family":"Cimuanga-Mukanya","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Faculty of Medicine, Pharmacy and Public Health, University of Mbujimayi, Mbujimayi P.O. Box 225, Congo"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evariste","family":"Tshibangu-Kabamba","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Faculty of Medicine, Pharmacy and Public Health, University of Mbujimayi, Mbujimayi P.O. Box 225, Congo"},{"name":"Department of Virology and Parasitology, Research Center for Infectious Disease Sciences, Graduate School of Medicine, Osaka Metropolitan University, Osaka 545-8585, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nadine","family":"Kayiba-Kalenda","sequence":"additional","affiliation":[{"name":"Department of Public Health, Faculty of Medicine, Pharmacy and Public Health, University of Mbujimayi, Mbujimayi P.O. Box 225, Congo"},{"name":"Research Institute of Health and Society, Catholic University of Louvain, 1200 Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tressy","family":"Kalenga-Ngomba","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Faculty of Medicine, University of Kinshasa, Kinshasa P.O. Box 834, Congo"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick de Jesus","family":"Ngoma-Kisoko","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Faculty of Medicine, University of Kinshasa, Kinshasa P.O. Box 834, Congo"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6938-8387","authenticated-orcid":false,"given":"Gunturu","family":"Revathi","sequence":"additional","affiliation":[{"name":"Department of Clinical Microbiology, Aga Khan University Hospital, Nairobi 00100, Kenya"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2663-9358","authenticated-orcid":false,"given":"Junko","family":"Akada","sequence":"additional","affiliation":[{"name":"Department of Environmental and Preventive Medicine, Faculty of Medicine, Oita University, Yufu 879-5593, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beno\u00eet","family":"Mbiya-Mukinayi","sequence":"additional","affiliation":[{"name":"Department of Paediatrics, Faculty of Medicine, Pharmacy and Public Health, University of Mbujimayi, Mbujimayi P.O. Box 225, Congo"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9519-5653","authenticated-orcid":false,"given":"Augustin Tshibaka","family":"Kabongo","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Faculty of Medicine, Pharmacy and Public Health, University of Mbujimayi, Mbujimayi P.O. Box 225, Congo"},{"name":"Department of Global Health, School of Tropical Medicine and Global Health, Nagasaki University, Nagasaki 852-8523, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ghislain","family":"Disashi-Tumba","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Faculty of Medicine, Pharmacy and Public Health, University of Mbujimayi, Mbujimayi P.O. Box 225, Congo"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6899-1686","authenticated-orcid":false,"given":"Takashi","family":"Matsumoto","sequence":"additional","affiliation":[{"name":"Department of Environmental and Preventive Medicine, Faculty of Medicine, Oita University, Yufu 879-5593, Japan"},{"name":"Research Center for Global and Local Infectious Diseases, Oita University, Yufu 879-5593, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1222-5819","authenticated-orcid":false,"given":"Yoshio","family":"Yamaoka","sequence":"additional","affiliation":[{"name":"Department of Environmental and Preventive Medicine, Faculty of Medicine, Oita University, Yufu 879-5593, Japan"},{"name":"Research Center for Global and Local Infectious Diseases, Oita University, Yufu 879-5593, Japan"},{"name":"Department of Medicine, Gastroenterology and Hepatology Section, Baylor College of Medicine, Houston, TX 77030, USA"},{"name":"Division of Gastroentero-Hepatology, Department of Internal Medicine, Faculty of Medicine-Dr. Soetomo Teaching Hospital, Universitas Airlangga, Surabaya 60286, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Ali, A., and AlHussaini, K.I. 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