pncA mutations in pyrazinamide-resistant Mycobacterium tuberculosis clinical isolates from the southeast region of Brazil

Barco, Patricia; Cardoso, Rosilene Fressatti; Hirata, Rosário Dc; Leite, Clarice Queico Fujimura; Pandolfi, J. R. C.; Sato, Daisy Nakamura; Shikama, M. L.; Melo, F. Fiuza de; Mamizuka, Elsa Masae; Campanerut, Paula Aline Zanetti; Hirata, Mário Hiroyuki

doi:10.1093/jac/dkl363

Cited by 48 publications

(46 citation statements)

References 41 publications

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“…Of 16 strains that were PZA resistant by the Wayne assay, 14 (87.5%) had mutations in the pncA gene, confirming that this is the principal mechanism for loss of PZase activity M. tuberculosis (2,8). Surprisingly, the two strains (no.…”

Section: Discussionmentioning

confidence: 92%

Evaluation of Colorimetric Methods Using Nicotinamide for Rapid Detection of Pyrazinamide Resistance in Mycobacterium tuberculosis

et al. 2010

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The direct detection of pyrazinamide resistance in Mycobacterium tuberculosis is sufficiently difficult that many laboratories do not attempt it. Most pyrazinamide resistance is caused by mutations that inactivate the pyrazinamidase enzyme needed to convert the prodrug pyrazinamide to its active form. We evaluated two newer and simpler methods to assess pyrazinamidase activity, the nitrate reductase and malachite green microtube assays, using nicotinamide in place of pyrazinamide. A total of 102 strains were tested by these methods and the results compared with those obtained by the classic Wayne assay. Mutations in the pncA gene were identified by sequencing the pncA genes from all isolates in which pyrazinamide resistance was detected by any of the three methods. Both the nitrate reductase and malachite green microtube assays showed sensitivities of 93.75% and specificities of 97.67%. Mutations in the pncA gene were found in 14 of 16 strains that were pyrazinamide resistant and in 1 of 4 strains that were sensitive by the Wayne assay. Both of these simple methods, used with nicotinamide, are promising and inexpensive alternatives for the rapid detection of pyrazinamide resistance in limited-resource countries.

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Section: Discussionmentioning

confidence: 92%

Evaluation of Colorimetric Methods Using Nicotinamide for Rapid Detection of Pyrazinamide Resistance in Mycobacterium tuberculosis

et al. 2010

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“…Furthermore, 5 (3.2%) PZA-susceptible isolates harbored 3 different mutations within pncA: Asp12Gly, Cys14Gly, and Ser59Phe. Although these mutations were observed among both PZA-resistant and PZAsusceptible isolates in previously published literature (10,15,26,27), of them, Asp12Gly was considered not to be associated with PZA resistance (20). The correlation of other two mutations with PZA resistance remains uncertain and deserves more exploration.…”

Section: Discussionmentioning

confidence: 94%

Phenotypic and Genotypic Characterization of Pyrazinamide Resistance among Multidrug-Resistant Mycobacterium tuberculosis Isolates in Zhejiang, China

Xia

Zhao

et al. 2015

Antimicrob Agents Chemother

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e To explore the phenotypic and genotypic characterization of pyrazinamide (PZA) resistance among multidrug-resistant Mycobacterium tuberculosis (MDR-TB) isolates in Zhejiang province, a total of 274 MDR-TB isolates were collected. Drug susceptibility testing and spoligotyping were performed on all clinical isolates. In addition, the mutated features of PZA-resistant loci, including pncA and rpsA, were also analyzed by DNA sequencing. Our results showed that the prevalence of PZA resistance among MDR-TB strains in Zhejiang province was 43.07% and that PZA resistance was associated with concomitant resistance to streptomycin. The majority of PZA-resistant MDR-TB isolates belonged to the Beijing family. Mutations within pncA, not rpsA, constituted the primary mechanism of PZA resistance. Among 118 PZA-resistant isolates, 53 different mutations were observed in pncA, and most of them were point mutations. Compared with the phenotypic data, DNA sequencing of pncA has sensitivity and specificity of 77.97% and 96.79%, respectively. Analysis of pncA provided a robust tool for rapid detection of PZA drug resistance.

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“…Correlations between genotypic clustering and pncA mutations were reported in studies from Brazil and Portugal (4,45,50). Strain clusters sharing the same pncA mutation in South Africa grouped together by spoligotype but differed by MIRU-VNTR type (41).…”

Section: Pyrazinamide-resistant Mycobacterium Tuberculosismentioning

confidence: 88%

“…Mutations in the putative promoter region have also been reported widely. The most common promoter mutation is Ϫ11 A ¡ G, which has been reported to occur in phylogenetically diverse strains (4,11,17,18,23,29,34,38,43,48,59). Other putative promoter mutations include Ϫ12 T ¡ C (41, 59), Ϫ10 T ¡ C (41), and Ϫ7 T ¡ C (43).…”

Section: Pyrazinamide-resistant Mycobacterium Tuberculosismentioning

confidence: 99%

“…SNPs on codon 57 are not uniquely restricted to M. bovis, as they may also occur in M. tuberculosis isolates (17,29). Alternative substitutions, such as 57 His ¡ Pro and 57 His ¡ Arg, are also possible (4,7,21). Another well-defined phylogenetic SNP in pncA is the synonymous substitution 46 Ala ¡ Ala in Mycobacterium canettii.…”

Section: Pyrazinamide-resistant Mycobacterium Tuberculosismentioning

confidence: 99%

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Systematic Analysis of Pyrazinamide-Resistant Spontaneous Mutants and Clinical Isolates of Mycobacterium tuberculosis

Stoffels

Mathys

Fauville-Dufaux

et al. 2012

Antimicrob Agents Chemother

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b Pyrazinamide (PZA) is a first-line antitubercular drug known for its activity against persistent Mycobacterium tuberculosis bacilli. We set out to systematically determine the PZA susceptibility profiles and mutations in the pyrazinamidase (pncA) gene of a collection of multidrug-resistant tuberculosis (MDR-TB) clinical isolates and PZA-resistant (PZA r ) spontaneous mutants. The frequency of acquired resistance to PZA was determined to be 10 ؊5 bacilli in vitro. Selection at a lower concentration of PZA yielded a significantly larger number of spontaneous mutants. The methodical approach employed allowed for determination of the frequency of the PZA r phenotype correlated with mutations in the pncA gene, which was 87.5% for the laboratory-selected spontaneous mutants examined in this study. As elucidated by structural analysis, most of the identified mutations were foreseen to affect protein activity through either alteration of an active site residue or destabilization of protein structure, indicating some preferential mutation site rather than random scattering. Twelve percent of the PZA r mutants did not have a pncA mutation, strongly indicating the presence of at least one other mechanism(s) of PZA r .

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pncA mutations in pyrazinamide-resistant Mycobacterium tuberculosis clinical isolates from the southeast region of Brazil

Abstract: These results contribute to the knowledge of the molecular mechanism of pyrazinamide resistance in Brazil and also expand the profile of pncA mutations worldwide. The MABA was successfully used to determine the MICs of pyrazinamide.

Cited by 48 publications

References 41 publications

Evaluation of Colorimetric Methods Using Nicotinamide for Rapid Detection of Pyrazinamide Resistance in Mycobacterium tuberculosis

Evaluation of Colorimetric Methods Using Nicotinamide for Rapid Detection of Pyrazinamide Resistance in Mycobacterium tuberculosis

Phenotypic and Genotypic Characterization of Pyrazinamide Resistance among Multidrug-Resistant Mycobacterium tuberculosis Isolates in Zhejiang, China

Systematic Analysis of Pyrazinamide-Resistant Spontaneous Mutants and Clinical Isolates of Mycobacterium tuberculosis

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