{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T13:54:46Z","timestamp":1791467686949,"version":"4.3.4"},"reference-count":35,"publisher":"American Society for Microbiology","license":[{"start":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T00:00:00Z","timestamp":1791417600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T00:00:00Z","timestamp":1791417600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100007280","name":"Srinakharinwirot University","doi-asserted-by":"publisher","award":["443\/2565, 064\/2566"],"award-info":[{"award-number":["443\/2565, 064\/2566"]}],"id":[{"id":"10.13039\/501100007280","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      The global spread of multidrug-resistant tuberculosis (MDR-TB) remains a major challenge to tuberculosis control programs, particularly in resource-limited settings where access to rapid molecular drug-susceptibility testing is limited. Resistance to isoniazid and rifampicin is most commonly associated with mutations in the\n                      <jats:italic toggle=\"yes\">katG<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">rpoB<\/jats:italic>\n                      genes, respectively, making these loci valuable molecular markers for early MDR-TB screening. In this study, we developed a dual loop-mediated isothermal amplification assay coupled with lateral flow detection for the simultaneous identification of the\n                      <jats:italic toggle=\"yes\">katG<\/jats:italic>\n                      S315T and\n                      <jats:italic toggle=\"yes\">rpoB<\/jats:italic>\n                      S531L mutations in\n                      <jats:italic toggle=\"yes\">Mycobacterium tuberculosis<\/jats:italic>\n                      . Mutation-specific primer sets were designed using mismatch amplification principles to achieve single-nucleotide specificity. Amplification was performed at 61\u00b0C for 60 min, followed by visual detection on a three-line lateral flow strip within 5 min. The assay demonstrated a preliminary detection threshold of approximately 10 CFU\/mL equivalents and exhibited no cross-reactivity with wild-type\n                      <jats:italic toggle=\"yes\">M. tuberculosis<\/jats:italic>\n                      H37Rv, non-tuberculous mycobacteria, or non-target resistance-associated mutations, including the\n                      <jats:italic toggle=\"yes\">inhA<\/jats:italic>\n                      \u221215 promoter mutation and\n                      <jats:italic toggle=\"yes\">rpoB<\/jats:italic>\n                      codon 526 variants. Preliminary clinical feasibility was evaluated using genetically characterized clinical sputum specimens and showed complete concordance with GenoType MTBDRplus and Sanger DNA sequencing results. Because the assay targets only two resistance-associated mutations, a negative result does not exclude isoniazid or rifampicin resistance mediated by alternative mutations.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      These proof-of-concept findings demonstrate the initial feasibility of a rapid, low-complexity molecular platform for the simultaneous detection of clinically important resistance-associated mutations in\n                      <jats:italic toggle=\"yes\">Mycobacterium tuberculosis<\/jats:italic>\n                      . The assay represents a promising step toward low-complexity molecular screening for selected multidrug-resistant tuberculosis-associated mutations, bringing us closer to simplified testing in settings where access to conventional molecular drug-resistance testing remains limited.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.02195-26","type":"journal-article","created":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T13:01:57Z","timestamp":1791464517000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Rapid simultaneous detection of\n                    <i>katG<\/i>\n                    S315T and\n                    <i>rpoB<\/i>\n                    S531L mutations for molecular screening of multidrug-resistant tuberculosis using a dual LAMP-lateral flow assay"],"prefix":"10.1128","author":[{"given":"Jutturong","family":"Ckumdee","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01qkghv97","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Clinical Pathology, Faculty of Medicine, Vajira Hospital, Navamindradhiraj University","place":["Bangkok, Thailand"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Somchai","family":"Santiwatanakul","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04718hx42","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Pathology, Faculty of Medicine, Srinakharinwirot University","place":["Nakhonnayok, Thailand"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nakarin","family":"Vutipow","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05gzceg21","id-type":"ROR","asserted-by":"publisher"}],"name":"Genetic Engineering and Bioinformatics Interdisciplinary Program, Graduate School, Kasetsart University","place":["Bangkok, Thailand"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0645-6450","authenticated-orcid":true,"given":"Thongchai","family":"Kaewphinit","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04718hx42","id-type":"ROR","asserted-by":"publisher"}],"name":"Center of Excellence in Medical and Environmental Innovation Research (CEMEIR), Srinakharinwirot University","place":["Bangkok, Thailand"]},{"id":[{"id":"https:\/\/ror.org\/04718hx42","id-type":"ROR","asserted-by":"publisher"}],"name":"Innovative Learning Center, Srinakharinwirot University","place":["Bangkok, Thailand"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","published-online":{"date-parts":[[2026,10,8]]},"reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(19)30308-3"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrdp.2016.76"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijid.2021.02.107"},{"key":"e_1_3_4_5_2","unstructured":"World Health Organization. 2025. Global tuberculosis report 2025. Geneva. 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