{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T07:02:38Z","timestamp":1779692558042,"version":"3.53.1"},"reference-count":31,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T00:00:00Z","timestamp":1779667200000},"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. Microbiol."],"abstract":"<jats:p>\n                    This study evaluated broth microdilution (BMD) and fluorescence PCR melting curve analysis (MeltPro MTB\/PZA, targeting\n                    <jats:italic>pncA<\/jats:italic>\n                    mutations) for detecting pyrazinamide (PZA) resistance-associated mutations in rifampicin-resistant tuberculosis (RR-TB). PZA susceptibility was assessed in RR-TB isolates from patients at TB prevention and control institutions and at designated hospitals in Beijing. Whole-genome sequencing (WGS) was used as a genotypic comparator to assess the agreement of BMD at critical concentrations (CCs) of 100 and 200\u202f\u03bcg\/mL and of the MeltPro assay. BMD showed resistance rates of 63.6% (70\/110) at 100\u202f\u03bcg\/mL and 45.5% (50\/110) at 200\u202f\u03bcg\/mL. Relative to WGS-defined genotypes (wild-type vs. mutation), BMD at 100\u202f\u03bcg\/mL showed 96.1% agreement for isolates with mutations and 64.4% for wild-type (\n                    <jats:italic>\u03ba<\/jats:italic>\n                    \u202f=\u202f0.590); at 200\u202f\u03bcg\/mL, agreement was 92.2% for mutations and 94.9% for wild-type (\n                    <jats:italic>\u03ba<\/jats:italic>\n                    \u202f=\u202f0.872). The MeltPro assay showed 89.7% agreement for isolates with mutations and 98.4% for wild-type (\n                    <jats:italic>\u03ba<\/jats:italic>\n                    \u202f=\u202f0.882). These findings indicate that using BMD at 200\u202f\u03bcg\/mL as the critical concentration achieves closer agreement with WGS-defined genotypes than 100\u202f\u03bcg\/mL, and that the MeltPro MTB\/PZA assay shows high concordance with WGS for rapid detection of pncA mutations in RR-TB isolates. Further validation using phenotypic reference methods and clinical outcomes is warranted.\n                  <\/jats:p>","DOI":"10.3389\/fmicb.2026.1828630","type":"journal-article","created":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T06:04:02Z","timestamp":1779689042000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Toward precision detection of pyrazinamide resistance: critical concentration assessment and rapid molecular method validation"],"prefix":"10.3389","volume":"17","author":[{"given":"Yanfeng","family":"Zhao","sequence":"first","affiliation":[{"name":"Beijing Center for Disease Prevention and Control","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lili","family":"Tian","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nenhan","family":"Wang","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Chen","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuangshuang","family":"Chen","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lan","family":"Yu","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengdi","family":"Pang","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beichuan","family":"Ding","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Li","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chuanyou","family":"Li","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaowei","family":"Dai","sequence":"additional","affiliation":[{"name":"Beijing Center for Disease Prevention and Control","place":["Beijing, China"]},{"name":"School of Public Health, Capital Medical University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,5,25]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"1164","DOI":"10.1111\/j.1469-0691.2009.03078.x","article-title":"Pyrazinamide resistance in multidrug-resistant Mycobacterium tuberculosis isolates in Japan","volume":"16","author":"Ando","year":"2010","journal-title":"Clin. 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