{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T18:09:48Z","timestamp":1790273388625,"version":"4.1.0"},"reference-count":73,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T00:00:00Z","timestamp":1790208000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"<jats:p>\n                    Effective control of tuberculosis, nontuberculous mycobacterial (NTM) disease, and melioidosis depends on rapid and accurate laboratory diagnosis.\n                    <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                    complex (MTBC) remains a health priority, whereas NTM infections, particularly\n                    <jats:italic>Mycobacterium avium<\/jats:italic>\n                    complex (MAC), are increasingly recognized and may be misdiagnosed as tuberculosis. Melioidosis, caused by\n                    <jats:italic>Burkholderia pseudomallei<\/jats:italic>\n                    , is underdiagnosed in endemic regions. These infections have overlapping clinical presentations, and conventional diagnostics methods may not accurately differentiate among these diseases. We developed and evaluated a quadruplex real-time PCR assay (TAM-mPCR) for simultaneous detection of MTBC, MAC, and\n                    <jats:italic>B. pseudomallei<\/jats:italic>\n                    , targeting\n                    <jats:italic>IS6110<\/jats:italic>\n                    ,\n                    <jats:italic>rrs<\/jats:italic>\n                    , and\n                    <jats:italic>BPSS1187<\/jats:italic>\n                    . A total of 607 pulmonary and extrapulmonary specimens from patients with suspected tuberculosis were collected at two sites in Thailand between 2024 and 2025. The limits of detection were 2 pg\/reaction for MTBC, 0.1 ng\/reaction for MAC, and 50 fg\/reaction for\n                    <jats:italic>B. pseudomallei<\/jats:italic>\n                    , with 100% analytical specificity. For MTBC detection, TAM-mPCR demonstrated 100% sensitivity (330\/330) and 99% (189\/191) specificity. All three culture-confirmed MAC specimens were detected. For\n                    <jats:italic>B. pseudomallei<\/jats:italic>\n                    , the assay demonstrated 73.2% sensitivity (30\/41) and 100% specificity (79\/79). For MTBC, TAM-mPCR showed high concordance with NAAT and culture results, but differed significantly from AFB microscopy (\n                    <jats:italic>P<\/jats:italic>\n                    \u2009&lt;\u20090.001). The assay also detected additional MTBC, MAC, and\n                    <jats:italic>B. pseudomallei<\/jats:italic>\n                    infections that were missed by routine diagnostics. TAM-mPCR enables rapid simultaneous pathogen detection and differentiation and may improve case detection and patient management in setting where tuberculosis and melioidosis co-endemic.\n                  <\/jats:p>","DOI":"10.1371\/journal.pntd.0014730","type":"journal-article","created":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T17:38:22Z","timestamp":1790271502000},"page":"e0014730","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and evaluation of a multiplex PCR for simultaneous detection of Mycobacterium tuberculosis complex, Mycobacterium avium complex, and Burkholderia pseudomallei in clinical specimens"],"prefix":"10.1371","volume":"20","author":[{"given":"Hasyanee","family":"Binmaeil","sequence":"first","affiliation":[{"name":"Mahidol University","place":["Bangkok, Thailand"],"department":["Department of Microbiology and Immunology, Faculty of Tropical Medicine"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thanakon","family":"Bunsong","sequence":"additional","affiliation":[{"name":"Mahidol University","place":["Bangkok, Thailand"],"department":["Department of Microbiology and Immunology, Faculty of Tropical Medicine"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Janjira","family":"Thaipadungpanit","sequence":"additional","affiliation":[{"name":"Mahidol University","place":["Bangkok, Thailand"],"department":["Department of Clinical Tropical Medicine, Faculty of Tropical Medicine"]},{"name":"Mahidol University","place":["Bangkok, Thailand"],"department":["Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wipawee","family":"Saenwongsa","sequence":"additional","affiliation":[{"name":"Ministry of Public Health","place":["Ubon Ratchathani, Thailand"],"department":["Office of Disease Prevention and Control Region 10th"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Montira","family":"Ngoenpramual","sequence":"additional","affiliation":[{"name":"Ministry of Public Health","place":["Ubon Ratchathani, Thailand"],"department":["Office of Disease Prevention and Control Region 10th"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sutthinan","family":"Chawsorngkorn","sequence":"additional","affiliation":[{"name":"Ministry of Public Health","place":["Ubon Ratchathani, Thailand"],"department":["Office of Disease Prevention and Control Region 10th"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soawarat","family":"Deekae","sequence":"additional","affiliation":[{"name":"Surin Hospital","place":["Surin, Thailand"],"department":["Department of Medical Technology and Clinical Pathology"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sunee","family":"Chayangsu","sequence":"additional","affiliation":[{"name":"Surin Hospital","place":["Surin, Thailand"],"department":["Department of Internal Medicine"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wang","family":"Nguitragool","sequence":"additional","affiliation":[{"name":"Mahidol University","place":["Bangkok, Thailand"],"department":["Department of Molecular Tropical Medicine and Genetics, Faculty of Tropical Medicine"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3906-7379","authenticated-orcid":true,"given":"Narisara","family":"Chantratita","sequence":"additional","affiliation":[{"name":"Mahidol University","place":["Bangkok, Thailand"],"department":["Department of Microbiology and Immunology, Faculty of Tropical Medicine"]},{"name":"Mahidol University","place":["Bangkok, Thailand"],"department":["Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine"]},{"name":"The Academy of Science, The Royal Society of Thailand","place":["Bangkok, Thailand"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,9,24]]},"reference":[{"issue":"1","key":"pntd.0014730.ref001","doi-asserted-by":"crossref","first-page":"15008","DOI":"10.1038\/nmicrobiol.2015.8","article-title":"Predicted global distribution of Burkholderia pseudomallei and burden of melioidosis","volume":"1","author":"D Limmathurotsakul","year":"2016","journal-title":"Nat Microbiol"},{"issue":"1","key":"pntd.0014730.ref002","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1055\/s-0034-1398389","article-title":"Melioidosis: Evolving concepts in epidemiology, pathogenesis, and treatment","volume":"36","author":"BJ Currie","year":"2015","journal-title":"Semin Respir Crit Care Med"},{"key":"pntd.0014730.ref003","doi-asserted-by":"crossref","first-page":"100118","DOI":"10.1016\/j.lansea.2022.100118","article-title":"Characteristics and One year outcomes of melioidosis patients in Northeastern Thailand: A 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