{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T13:09:25Z","timestamp":1789736965352,"version":"4.0.1"},"reference-count":28,"publisher":"American Society for Microbiology","license":[{"start":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:00:00Z","timestamp":1789689600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:00:00Z","timestamp":1789689600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"Special Health Fund for Mass Immunization and Disease Control","award":["024.00129238\/2024-14"],"award-info":[{"award-number":["024.00129238\/2024-14"]}]}],"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                      Drug-resistant tuberculosis (TB) remains a major global health challenge, particularly in high-burden settings, where rapid detection of resistance to rifampicin (RIF) and isoniazid (INH) is essential for timely treatment. The FluoroType MTBDR assay is an automated real-time PCR platform for simultaneous detection of\n                      <jats:italic toggle=\"yes\">Mycobacterium tuberculosis<\/jats:italic>\n                      complex (MTBC) and resistance to RIF and INH. This study aimed to evaluate the performance of FluoroType using clinical MTBC isolates under routine laboratory conditions in a reference setting. The assay was prospectively assessed in comparison with the GenoType MTBDR\n                      <jats:italic toggle=\"yes\">plus<\/jats:italic>\n                      line probe assay (LPA) using 1,841 clinical isolates, with discordant results resolved by Sanger sequencing. For MTBC detection, FluoroType achieved 100% sensitivity (1,807\/1,807), 88.2% specificity (30\/34), and 99.8% overall accuracy. FluoroType demonstrated 99.9% (1,683\/1,685) sensitivity in detecting wild-type isolates. For resistance detection, FluoroType showed 95.5% sensitivity for RIF (42\/44) and 94.4% for INH (34\/36). Among multidrug-resistant isolates, sensitivity was 85.4% (35\/41). Discordances were mainly associated with heteroresistance, and uncommon or synonymous mutations. Importantly, limitations in identifying specific mutations highlight the need for cautious interpretation and, when necessary, confirmatory sequencing. Overall, these findings support FluoroType as a rapid, high-throughput, and reliable tool for MTBC detection and drug resistance profiling in reference laboratories.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      This study is the first to prospectively evaluate the FluoroType MTBDR assay performance using a large collection of clinical isolates under routine conditions in a high-burden tuberculosis (TB) reference laboratory. It assesses detection of\n                      <jats:italic toggle=\"yes\">Mycobacterium tuberculosis<\/jats:italic>\n                      complex and resistance to rifampicin and isoniazid. Drug-resistant TB remains a major global health challenge, requiring rapid and reliable diagnostic tools. FluoroType demonstrated high accuracy and operational advantages over the line probe assay (LPA), including a streamlined workflow and reduced turnaround time for high-throughput settings. Importantly, this study highlights limitations shared by molecular assays, particularly in detecting and interpreting heteroresistance, uncommon mutations, and synonymous rpoB variants, which may lead to discrepant resistance classification across platforms. These findings underscore the need for cautious interpretation of molecular results and sequencing to resolve clinically relevant discrepancies. Overall, FluoroType is a valuable addition to TB diagnostic algorithms, but results should be interpreted in the context of assay-specific limitations.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.01880-26","type":"journal-article","created":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T13:00:16Z","timestamp":1789736416000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Real-world evaluation of the FluoroType MTBDR 2.0 assay for detection of rifampicin and isoniazid resistance-conferring mutations in\n                    <i>Mycobacterium tuberculosis<\/i>\n                    complex clinical isolates"],"prefix":"10.1128","author":[{"given":"Fabiane Maria de Almeida","family":"Ferreira","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02wna9e57","id-type":"ROR","asserted-by":"publisher"}],"name":"N\u00facleo de Tuberculose e Micobacterioses, Centro de Bacteriologia, Instituto Adolfo Lutz (IAL)","place":["S\u00e3o Paulo, Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angela Pires","family":"Brand\u00e3o","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02wna9e57","id-type":"ROR","asserted-by":"publisher"}],"name":"N\u00facleo de Tuberculose e Micobacterioses, Centro de Bacteriologia, Instituto Adolfo Lutz (IAL)","place":["S\u00e3o Paulo, Brazil"]},{"id":[{"id":"https:\/\/ror.org\/04jhswv08","id-type":"ROR","asserted-by":"publisher"}],"name":"Instituto Oswaldo Cruz, Funda\u00e7\u00e3o Oswaldo Cruz (FIOCRUZ)","place":["Rio de Janeiro, Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thiago Brobio","family":"Massanti","sequence":"additional","affiliation":[{"name":"Mobius Lifescience","place":["Pinhais, Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonio Carlos da Silva","family":"Rego","sequence":"additional","affiliation":[{"name":"Mobius Lifescience","place":["Pinhais, Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucilaine","family":"Ferrazoli","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02wna9e57","id-type":"ROR","asserted-by":"publisher"}],"name":"N\u00facleo de Tuberculose e Micobacterioses, Centro de Bacteriologia, Instituto Adolfo Lutz (IAL)","place":["S\u00e3o Paulo, Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erica","family":"Chimara","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02wna9e57","id-type":"ROR","asserted-by":"publisher"}],"name":"N\u00facleo de Tuberculose e Micobacterioses, Centro de Bacteriologia, Instituto Adolfo Lutz (IAL)","place":["S\u00e3o Paulo, Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ros\u00e2ngela Siqueira","family":"de Oliveira","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02wna9e57","id-type":"ROR","asserted-by":"publisher"}],"name":"N\u00facleo de Tuberculose e Micobacterioses, Centro de Bacteriologia, Instituto Adolfo Lutz (IAL)","place":["S\u00e3o Paulo, Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5758-6688","authenticated-orcid":true,"given":"Juliana Maira Watanabe","family":"Pinhata","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02wna9e57","id-type":"ROR","asserted-by":"publisher"}],"name":"N\u00facleo de Tuberculose e Micobacterioses, Centro de Bacteriologia, Instituto Adolfo Lutz (IAL)","place":["S\u00e3o Paulo, Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","published-online":{"date-parts":[[2026,9,18]]},"reference":[{"key":"e_1_3_4_2_2","unstructured":"World Health Organization. 2025. Global tuberculosis report 2025. World Health Organization. Geneva Switzerland"},{"key":"e_1_3_4_3_2","unstructured":"Minist\u00e9rio da Sa\u00fade. 2026. Boletim epidemiol\u00f3gico tuberculose 2026. Minist\u00e9rio Da Sa\u00fade. Bras\u00edlia Brazil"},{"key":"e_1_3_4_4_2","doi-asserted-by":"crossref","unstructured":"Migliori GB Bothamley G Duarte R Rendon A eds. 2018. Tuberculosis. European Respiratory Society. Sheffield United Kingdom","DOI":"10.1183\/2312508X.erm8218"},{"key":"e_1_3_4_5_2","unstructured":"K\u00f6dm\u00f6n C. 2018. Handbook on tuberculosis laboratory diagnostic methods in the European Union. European Centre for Disease Prevention and Control. Stockholm Sweden"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.medmal.2017.04.008"},{"key":"e_1_3_4_7_2","unstructured":"World Health Organization. 2025. WHO consolidated guidelines on tuberculosis: module 3: diagnosis. World Health Organization. Geneva Switzerland"},{"key":"e_1_3_4_8_2","unstructured":"Hain Lifescience GmbH. 2017. FluoroLyse VER 1.0: kit for manual extraction of bacterial DNA from patient specimens and cultivated samples. Hain Lifescience GmbH. Nehren Germany"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00531-18"},{"key":"e_1_3_4_10_2","unstructured":"World Health Organization. 2019. WHO evaluation of centralized assays for detection of tuberculosis and resistance to rifampicin and isoniazid. World Health Organization. Geneva Switzerland"},{"key":"e_1_3_4_11_2","unstructured":"World Health Organization. 2021. WHO consolidated guidelines on tuberculosis: module 3: diagnosis. rapid diagnostics for tuberculosis detection. World Health Organization. Geneva Switzerland"},{"key":"e_1_3_4_12_2","first-page":"9","article-title":"Practical issues in calculating the sample size for prevalence studies","volume":"1","author":"Naing L","year":"2006","unstructured":"Naing L, Winn T, Rusli BN. 2006. Practical issues in calculating the sample size for prevalence studies. Arch Orofac Sci 1:9\u201314.","journal-title":"Arch Orofac Sci"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.000734"},{"key":"e_1_3_4_14_2","unstructured":"Hain Lifescience GmbH. 2015. GenoType MTBDRplus VER 2.0: molecular genetic assay for identification of the Mycobacterium tuberculosis complex and its resistance to rifampicin and isoniazid from clinical specimens and cultivated samples. Hain Lifescience GmbH. Nehren Germany"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2180-8-48"},{"key":"e_1_3_4_16_2","unstructured":"Hain Lifescience GmbH. 2021. FluoroType MTBDR VER 2.0: molecular genetic assay for automated detection of the Mycobacterium tuberculosis complex and its resistance to rifampicin and isoniazid from sputum specimens and cultivated samples. Hain Lifescience GmbH. Nehren Germany"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10096-021-04316-0"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1089\/mdr.2008.0798"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkt090"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.37.9.3022-3024.1999"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1590\/1806-3713\/e20180128"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmi.2021.04.022"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmi.2020.12.020"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.5588\/ijtld.25.0150"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tube.2020.102004"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1590\/0037-8682-0184-2025"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00364-21"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00812-17"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.00912-25"}],"container-title":["Microbiology Spectrum"],"original-title":[],"contributor":[{"given":"Sydney","family":"Stanley","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"reviewer"}]}],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.01880-26","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.01880-26","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T13:00:21Z","timestamp":1789736421000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/spectrum.01880-26"}},"subtitle":[],"editor":[{"given":"Kenneth","family":"Gavina","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,9,18]]},"references-count":28,"alternative-id":["10.1128\/spectrum.01880-26"],"URL":"https:\/\/doi.org\/10.1128\/spectrum.01880-26","relation":{},"ISSN":["2165-0497"],"issn-type":[{"value":"2165-0497","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,18]]},"assertion":[{"value":"2026-06-05","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-08-14","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-18","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e01880-26"}}