{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T21:44:58Z","timestamp":1788471898491,"version":"build-2803163510"},"reference-count":32,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T00:00:00Z","timestamp":1778716800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      The reliability of phenotypic pyrazinamide (PZA) drug susceptibility test (DST) in\n                      <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                      complex (MTBC) isolates is under growing scrutiny. The MGIT 960 System, the only WHO-endorsed method, has shown increasing false positive (FP) rates. In 2024, we reported a surge, and in 2025 Becton, Dickinson, and Company (BD) issued a global alert and suspended the PZA kit production. We assessed the extent of FP PZA resistance (PZA-R), its diagnostic impact, and the ways to distinguish FP from true positive (TP) results in our laboratory.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Between 2023 and 2025, 9% of the isolates were confirmed as TP PZA-R, mostly due to\n                      <jats:italic>M. bovis<\/jats:italic>\n                      . The MGIT 960 FP rate rose from 18% in 2023 to 33% in early 2025. Among 176 isolates, 47 (27%) were FP, 16 (9%) were TP, and 112 (64%) were true negatives (TN). The TP isolates had a longer TTP than the FP isolates (7.2 vs 5.7 days). The FP isolates showed MGIT intermediate growth index values between TP and TN (\n                      <jats:italic>p<\/jats:italic>\n                      &amp;lt; 0.005). Notably, the H37Rv reference strain had a 25% FP rate. The confirmatory testing increased the workload but prevented misreporting.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      Our data reveal a high FP rate for PZA-R with the MGIT 960 System, with no reliable phenotypic alternative available. The sequencing of\n                      <jats:italic>pncA<\/jats:italic>\n                      has been crucial in preventing misreporting and ensuring accurate TB treatment decisions. There is an urgent need for better phenotypic tools and broader molecular approaches beyond\n                      <jats:italic>pncA<\/jats:italic>\n                      so that PZA-R can be detected more reliably and on a scale that supports everyday clinical practice.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fcimb.2026.1777714","type":"journal-article","created":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T05:41:24Z","timestamp":1778737284000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Pyrazinamide susceptibility testing in Mycobacterium tuberculosis: impact of false-positive BD MGIT 960 results and added value of sequencing for accurate diagnosis in a low-MDR-TB-incidence context"],"prefix":"10.3389","volume":"16","author":[{"given":"Touyana","family":"Semenova","sequence":"first","affiliation":[{"name":"Institute of Microbiology, Lausanne University and Lausanne University Hospital","place":["Lausanne, Switzerland"]},{"name":"Microbiology Laboratory, Unilabs","place":["Coppet, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gr\u00e9gory","family":"Gonzalez","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Lausanne University and Lausanne University Hospital","place":["Lausanne, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beno\u00eet","family":"Lechartier","sequence":"additional","affiliation":[{"name":"Division of Pulmonology, Department of Medicine, Lausanne University and Lausanne University Hospital (CHUV)","place":["Lausanne, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Sander","sequence":"additional","affiliation":[{"name":"Institute of Medical Microbiology, University of Zurich","place":["Zurich, Switzerland"]},{"name":"National Reference Center for Mycobacteria","place":["Zurich, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christophe","family":"von Garnier","sequence":"additional","affiliation":[{"name":"Division of Pulmonology, Department of Medicine, Lausanne University and Lausanne University Hospital (CHUV)","place":["Lausanne, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bettina","family":"Schulthess","sequence":"additional","affiliation":[{"name":"Institute of Medical Microbiology, University of Zurich","place":["Zurich, Switzerland"]},{"name":"National Reference Center for Mycobacteria","place":["Zurich, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jesica","family":"Mazza-Stalder","sequence":"additional","affiliation":[{"name":"Division of Pulmonology, Department of Medicine, Lausanne University and Lausanne University Hospital (CHUV)","place":["Lausanne, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gilbert","family":"Greub","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Lausanne University and Lausanne University Hospital","place":["Lausanne, Switzerland"]},{"name":"Infectious Disease Service, Lausanne University and Lausanne University Hospital","place":["Lausanne, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Onya","family":"Opota","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Lausanne University and Lausanne University Hospital","place":["Lausanne, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,5,14]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.meegid.2013.06.008","article-title":"Mycobacterium tuberculosis isolates from Rio de Janeiro reveal unusually low correlation between pyrazinamide resistance and mutations in the pncA gene","volume":"19","author":"Bhuju","year":"2013","journal-title":"Infect. Genet. Evol."},{"key":"B2","doi-asserted-by":"publisher","first-page":"600","DOI":"10.1128\/AAC.24.4.600","article-title":"Susceptibility of Mycobacterium tuberculosis to pyrazinamide and its relationship to pyrazinamidase activity","volume":"24","author":"Butler","year":"1983","journal-title":"Antimicrob. Agents Chemother."},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1093\/jacamr\/dlae037","article-title":"Prediction of pyrazinamide resistance in Mycobacterium tuberculosis using structure-based machine-learning approaches","volume":"6","author":"Carter","year":"2024","journal-title":"JAC-Antimicrob. Resist."},{"key":"B4","doi-asserted-by":"publisher","first-page":"300","DOI":"10.1128\/JCM.01775-09","article-title":"Potential for erroneous results indicating resistance when using the Bactec MGIT 960 system for testing susceptibility of Mycobacterium tuberculosis to pyrazinamide","volume":"48","author":"Chedore","year":"2010","journal-title":"J. Clin. Microbiol."},{"key":"B5","doi-asserted-by":"publisher","first-page":"789","DOI":"10.1128\/AAC.01876-12","article-title":"A time-to-event pharmacodynamic model describing treatment response in patients with pulmonary tuberculosis using days to positivity in automated liquid mycobacterial culture","volume":"57","author":"Chigutsa","year":"2013","journal-title":"Antimicrob. Agents Chemother."},{"key":"B6","doi-asserted-by":"publisher","first-page":"537","DOI":"10.1038\/31159","article-title":"Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence","volume":"393","author":"Cole","year":"1998","journal-title":"Nature"},{"key":"B7","doi-asserted-by":"publisher","first-page":"1374","DOI":"10.1128\/JCM.03197-12","article-title":"Evaluation of methods for testing the susceptibility of clinical Mycobacterium tuberculosis isolates to pyrazinamide","volume":"51","author":"Cui","year":"2013","journal-title":"J. Clin. Microbiol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.tube.2011.05.006","article-title":"Pyrazinamide pharmacokinetics and efficacy in adults and children","volume":"92","author":"Donald","year":"2012","journal-title":"Tuberculosis. (Edinb)."},{"key":"B9","doi-asserted-by":"publisher","first-page":"121.e1","DOI":"10.1016\/j.cmi.2024.09.024","article-title":"Evaluating the efficacy of whole genome sequencing in predicting susceptibility profiles for first-line antituberculosis drugs","volume":"31","author":"Ghodousi","year":"2025","journal-title":"Clin. Microbiol. Infect."},{"key":"B10","doi-asserted-by":"publisher","first-page":"1852","DOI":"10.1128\/AAC.00110-08","article-title":"Pyrazinamide resistance and pncA gene mutations in Mycobacterium tuberculosis","volume":"52","author":"Jur\u00e9en","year":"2008","journal-title":"Antimicrob. Agents Chemother."},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1128\/MMBR.00070-19","article-title":"The bewildering antitubercular action of pyrazinamide","volume":"84","author":"Lamont","year":"2020","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"B12","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1128\/AAC.01209-13","article-title":"Sensititre MYCOTB MIC plate for testing Mycobacterium tuberculosis susceptibility to first- and second-line drugs","volume":"58","author":"Lee","year":"2014","journal-title":"Antimicrob. Agents Chemother."},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2020.594171","article-title":"Characterization of pncA mutations and prediction of PZA resistance in Mycobacterium tuberculosis clinical isolates from Chongqing, China","volume":"11","author":"Li","year":"2021","journal-title":"Front. Microbiol."},{"key":"B14","doi-asserted-by":"publisher","first-page":"556","DOI":"10.1128\/AAC.20.4.556","article-title":"Use of pyrazinamidase activity on Mycobacterium tuberculosis as a rapid method for determination of pyrazinamide susceptibility","volume":"20","author":"McClatchy","year":"1981","journal-title":"Antimicrob. Agents Chemother."},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01819-14","article-title":"Mycobacterium tuberculosis pyrazinamide resistance determinants: a multicenter study","volume":"5","author":"Miotto","year":"2014","journal-title":"mBio"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.02617-20","article-title":"Overcoming the challenges of pyrazinamide susceptibility testing in clinical Mycobacterium tuberculosis isolates","volume":"65","author":"Mok","year":"2021","journal-title":"Antimicrob. Agents Chemother."},{"key":"B17","doi-asserted-by":"publisher","first-page":"3748163","DOI":"10.1155\/2017\/3748163","article-title":"Using reduced inoculum densities of Mycobacterium tuberculosis in MGIT pyrazinamide susceptibility testing to prevent false-resistant results and improve accuracy: a multicenter evaluation","author":"Morlock","year":"2017","journal-title":"Tuberc. Res. Treat."},{"key":"B18","doi-asserted-by":"publisher","first-page":"3552","DOI":"10.1128\/JCM.01437-17","article-title":"Improved Bactec MGIT 960 pyrazinamide test decreases detection of false Mycobacterium tuberculosis pyrazinamide resistance","volume":"55","author":"Mustazzolu","year":"2017","journal-title":"J. Clin. Microbiol."},{"key":"B19","doi-asserted-by":"publisher","first-page":"291","DOI":"10.1128\/JCM.01838-12","article-title":"Prevention of false resistance results obtained in testing the susceptibility of Mycobacterium tuberculosis to pyrazinamide with the Bactec MGIT 960 system using a reduced inoculum","volume":"51","author":"Piersimoni","year":"2013","journal-title":"J. Clin. Microbiol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"27091","DOI":"10.1038\/s41598-024-77947-w","article-title":"Leveraging large-scale Mycobacterium tuberculosis whole genome sequence data to characterise drug-resistant mutations using machine learning and statistical approaches","volume":"14","author":"Pruthi","year":"2024","journal-title":"Sci. Rep."},{"key":"B21","doi-asserted-by":"publisher","first-page":"5267","DOI":"10.1128\/AAC.00204-15","article-title":"Systematic review of mutations in pyrazinamidase associated with pyrazinamide resistance in Mycobacterium tuberculosis clinical isolates","volume":"59","author":"Ramirez-Busby","year":"2015","journal-title":"Antimicrob. Agents Chemother."},{"key":"B22","first-page":"62","article-title":"Genomic polymorphisms in a laboratory isolate of Mycobacterium tuberculosis reference strain H37Rv (ATCC27294)","volume":"40","author":"Santoro","year":"2017","journal-title":"New Microbiol."},{"key":"B23","doi-asserted-by":"publisher","first-page":"662","DOI":"10.1038\/nm0696-662","article-title":"Mutations in pncA, a gene encoding pyrazinamidase\/nicotinamidase, cause resistance to the antituberculous drug pyrazinamide in tubercle bacillus","volume":"2","author":"Scorpio","year":"1996","journal-title":"Nat. Med."},{"key":"B24","doi-asserted-by":"publisher","first-page":"769","DOI":"10.1186\/s12864-017-4146-z","article-title":"A multiple genome analysis of Mycobacterium tuberculosis reveals specific novel genes and mutations associated with pyrazinamide resistance","volume":"18","author":"Sheen","year":"2017","journal-title":"BMC Genomics"},{"key":"B25","doi-asserted-by":"publisher","first-page":"4985","DOI":"10.2147\/IDR.S368444","article-title":"Pyrazinamide resistance and pncA mutation profiles in multidrug resistant Mycobacterium tuberculosis","volume":"15","author":"Shi","year":"2022","journal-title":"Infect. Drug Resist."},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00145-19","article-title":"Direct detection of pyrazinamide resistance in Mycobacterium tuberculosis by use of pncA PCR sequencing","volume":"57","author":"Tam","year":"2019","journal-title":"J. Clin. Microbiol."},{"key":"B27","volume-title":"Technical Manual for Drug Susceptibility Testing of Medicines Used in the Treatment of Tuberculosis","year":"2018"},{"key":"B28","volume-title":"Catalogue of Mutations in Mycobacterium Tuberculosis Complex and their Association with Drug Resistance","year":"2023"},{"key":"B29","doi-asserted-by":"publisher","first-page":"147","DOI":"10.1016\/j.meegid.2018.11.012","article-title":"pncA gene mutations in reporting pyrazinamide resistance among the MDR-TB suspects","volume":"72","author":"Wu","year":"2019","journal-title":"Infect. Genet. Evol."},{"key":"B30","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1099\/0022-1317-51-1-42","article-title":"Conditions that may affect the results of susceptibility testing of Mycobacterium tuberculosis to pyrazinamide","volume":"51","author":"Zhang","year":"2002","journal-title":"J. Med. Microbiol."},{"key":"B31","doi-asserted-by":"publisher","first-page":"2044","DOI":"10.1128\/JB.181.7.2044-2049.1999","article-title":"Role of acid pH and deficient efflux of pyrazinoic acid in unique susceptibility of Mycobacterium tuberculosis to pyrazinamide","volume":"181","author":"Zhang","year":"1999","journal-title":"J. Bacteriol."},{"key":"B32","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1128\/microbiolspec.MGM2-0023-2013","article-title":"Mechanisms of pyrazinamide action and resistance","volume":"2","author":"Zhang","year":"2013","journal-title":"Microbiol. Spectr."}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2026.1777714\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T05:41:31Z","timestamp":1778737291000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2026.1777714\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,14]]},"references-count":32,"alternative-id":["10.3389\/fcimb.2026.1777714"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2026.1777714","relation":{},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,14]]},"article-number":"1777714"}}