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Detection of Novel Gene Mutations Associated with Pyrazinamide Resistance in Multidrug-Resistant Clinical Isolates in Southern China. | LitMetric
Objective : Pyrazinamide (PZA) is a cornerstone of modern tuberculosis regimens. This study aimed to investigate the performance of genotypic testing of upstream region, and genes to add insights for more accurate molecular diagnosis of PZA-resistant (R) Methods : Drug susceptibility testing, sequencing analysis of PZA-related genes including the entire operon of () and PZase assay were performed for 448 clinical isolates.Results : Our data showed that among 448 clinical isolates, 113 were MDR, 195 pre-XDR and 70 XDR TB, while the remaining 70 strains had other combinations of drug-resistance. A total of 60.04% (269/448) clinical isolates were resistant to PZA, of which 78/113 were MDR, 119/195 pre-XDR and 29/70 XDR TB strains. PZA isolates have predominance (83.3%) of Beijing genotype. Genotypic characterization of revealed novel nonsynonymous mutations in with negative PZase activity which led to confer PZA. Compared with phenotypic data, 84.38% (227/269) PZA strains with mutations in upstream region exhibited 83.64% sensitivity but the combined evaluation of the mutations in 2.60% (7/269), 1.48% (4/269), 1.11% (3/269) and 0.74% (2/269) increased the sensitivity to 89.59%. Fifty-seven novel mutations were identified in this study. Interestingly, a frameshift deletion (C-114del) in upstream of nullified the effect of A-11G mutation and induced positive PZase activity, divergent from five PZase negative A-11G PZA mutants. Twenty-six PZA strains having wild-type-sequenced genes with positive or negative PZase suggest the existence of unknown resistance mechanisms.Conclusion : Our study revealed that PZA rate in MDR and pre-XDR TB was markedly higher in southern China. The concomitant evaluation of UFR, and provides more dependable genotypic results of PZA resistance. Fifty-seven novel mutations/indels in this study may play a vital role as diagnostic markers. The upstream region of and PZase regulation are valuable to explore the unknown mechanism of PZA-resistance.
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