{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,30]],"date-time":"2026-08-30T00:40:44Z","timestamp":1788050444043,"version":"build-2784847793"},"reference-count":34,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2025,7,9]],"date-time":"2025-07-09T00:00:00Z","timestamp":1752019200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,7,9]],"date-time":"2025-07-09T00:00:00Z","timestamp":1752019200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82472368"],"award-info":[{"award-number":["82472368"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Bureau of Science and Information Technology of Guangzhou Municipality | Guangzhou Municipal Science and Technology Project","award":["2023A03J0991"],"award-info":[{"award-number":["2023A03J0991"]}]},{"DOI":"10.13039\/501100005270","name":"Fujian Provincial Department of Science and Technology","doi-asserted-by":"publisher","award":["2023D012"],"award-info":[{"award-number":["2023D012"]}],"id":[{"id":"10.13039\/501100005270","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100016078","name":"Fuzhou Science and Technology Bureau","doi-asserted-by":"publisher","award":["2023-S-042"],"award-info":[{"award-number":["2023-S-042"]}],"id":[{"id":"10.13039\/100016078","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010166","name":"Xiamen Municipal Bureau of Science and Technology","doi-asserted-by":"publisher","award":["502Z20231034"],"award-info":[{"award-number":["502Z20231034"]}],"id":[{"id":"10.13039\/100010166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2025,7,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Fluoroquinolones (FQs) resistance in\n                      <jats:italic toggle=\"yes\">Mycobacterium tuberculosis<\/jats:italic>\n                      (MTB), primarily driven by mutations in the quinolone resistance-determining region (QRDR) of\n                      <jats:italic toggle=\"yes\">gyrA<\/jats:italic>\n                      , poses a major concern in treating for multidrug-resistant tuberculosis (MDR-TB). These QRDR mutations are known to confer varying levels of resistance, leading to differences in treatment outcomes. Here, we introduced the MeltArray MTB\/FQs assay, a multiplex PCR method that detected 11\n                      <jats:italic toggle=\"yes\">gyrA<\/jats:italic>\n                      -QRDR mutations and quantified their fractions via a polynomial regression algorithm-based formula, enabling mutation identification and quantification within 3 h in one reaction. This assay, with a limit of detection (LOD) of 50 copies\/reaction, could identify mixtures containing 5% mutant gDNA across 50 to 50,000 copies\/reaction. Clinical evaluation of 442 culture samples displayed 95.23% sensitivity and 99.32% specificity compared with phenotypic antimicrobial susceptibility testing (pAST). Evaluation of 121 paired sputum-culture samples revealed sensitivities of 90.32% in sputum samples and 95.24% in culture samples, both with specificities of 100%, when compared with pAST. Further evaluation of 285 sputum samples showed 93.75% positive percent agreement (PPA) and 98.10% negative percent agreement (NPA) in comparison with the MeltPro MTB\/FQs kit (Zeesan Biotech, China). All mutant samples identified by MeltArray MTB\/FQs but classified as susceptible by pAST or as wild type by MeltPro MTB\/FQs were confirmed through Sanger sequencing and droplet digital PCR (ddPCR). The formula for predicting mutation fraction (MUT%) showed accuracy rates of 88.00%, 88.89%, and 83.33% in the training, validation, and test data sets, respectively, when compared with ddPCR results. Overall, MeltArray MTB\/FQs assay offers an upgraded alternative for routine FQs resistance monitoring.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Rising FQs resistance has driven the spread of pre-extensively drug-resistant tuberculosis (pre-XDR-TB), challenging global tuberculosis (TB) control efforts. Conventional molecular assays for FQs resistance often cannot distinguish between low-level and high-level resistance mutations or detect low-fraction heteroresistant populations. In this study, we established a MeltArray MTB\/FQs assay that can identify all the 11 critical mutations in the\n                        <jats:italic toggle=\"yes\">gyrA<\/jats:italic>\n                        -QRDR with a LOD of 50 copies\/reaction, enabling direct, culture-independent analysis of sputum samples. By using an algorithm to quantify mutations at levels as low as 5% in mixtures, MeltArray achieved both mutation identification and quantification within 3 h in a reaction, thus providing a powerful tool for early detection and precise management of pre-XDR-TB.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/jcm.00146-25","type":"journal-article","created":{"date-parts":[[2025,6,3]],"date-time":"2025-06-03T09:00:39Z","timestamp":1748941239000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":2,"title":["Identification and quantification of\n                    <i>gyrA<\/i>\n                    variants in fluoroquinolone-resistant\n                    <i>Mycobacterium tuberculosis<\/i>\n                    in a MeltArray reaction"],"prefix":"10.1128","volume":"63","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-8829-7180","authenticated-orcid":false,"given":"Chunxia","family":"Tu","sequence":"first","affiliation":[{"name":"Engineering Research Centre of Molecular Diagnostics of the Ministry of Education, State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University","place":["Xiamen, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Biyi","family":"Su","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04szr1369","id-type":"ROR","asserted-by":"publisher"}],"name":"Guangzhou Chest Hospital","place":["Guangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunfan","family":"Xiong","sequence":"additional","affiliation":[{"name":"Engineering Research Centre of Molecular Diagnostics of the Ministry of Education, State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University","place":["Xiamen, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zihan","family":"Xia","sequence":"additional","affiliation":[{"name":"Engineering Research Centre of Molecular Diagnostics of the Ministry of Education, State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University","place":["Xiamen, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunlin","family":"Xiang","sequence":"additional","affiliation":[{"name":"Engineering Research Centre of Molecular Diagnostics of the Ministry of Education, State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University","place":["Xiamen, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5219-1265","authenticated-orcid":true,"given":"Yaoju","family":"Tan","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04szr1369","id-type":"ROR","asserted-by":"publisher"}],"name":"Guangzhou Chest Hospital","place":["Guangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5066-4698","authenticated-orcid":false,"given":"Ye","family":"Xu","sequence":"additional","affiliation":[{"name":"Engineering Research Centre of Molecular Diagnostics of the Ministry of Education, State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University","place":["Xiamen, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3431-138X","authenticated-orcid":true,"given":"Qingge","family":"Li","sequence":"additional","affiliation":[{"name":"Engineering Research Centre of Molecular Diagnostics of the Ministry of Education, State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University","place":["Xiamen, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1407426"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.02516-18"},{"key":"e_1_3_3_4_2","volume-title":"Global Tuberculosis Report 2024","year":"2024","unstructured":"2024. 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