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Evaluation of colorimetric methods using nicotinamide for rapid detection of pyrazinamide resistance in Mycobacterium tuberculosis

(2010) JOURNAL OF CLINICAL MICROBIOLOGY. 48(8). p.2729-2733
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Abstract
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.
Keywords
TUBERCLE-BACILLI, DRUG-RESISTANCE, ALTERNATIVE METHODS, RIFAMPICIN RESISTANCE, NITRATE REDUCTASE ASSAY, TESTING SUSCEPTIBILITY, PNCA MUTATIONS, PYRAZINOIC ACID, STRAINS, SYSTEM

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Citation

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Chicago
Mirabal, Niuris C, Sergio L Yzquierdo, Dihadenys Lemus, Mariela Madruga, Yoslaine Milian, Miguel Echemendia, Howard Takiff, et al. 2010. “Evaluation of Colorimetric Methods Using Nicotinamide for Rapid Detection of Pyrazinamide Resistance in Mycobacterium Tuberculosis.” Journal of Clinical Microbiology 48 (8): 2729–2733.
APA
Mirabal, N. C., Yzquierdo, S. L., Lemus, D., Madruga, M., Milian, Y., Echemendia, M., Takiff, H., et al. (2010). Evaluation of colorimetric methods using nicotinamide for rapid detection of pyrazinamide resistance in Mycobacterium tuberculosis. JOURNAL OF CLINICAL MICROBIOLOGY, 48(8), 2729–2733.
Vancouver
1.
Mirabal NC, Yzquierdo SL, Lemus D, Madruga M, Milian Y, Echemendia M, et al. Evaluation of colorimetric methods using nicotinamide for rapid detection of pyrazinamide resistance in Mycobacterium tuberculosis. JOURNAL OF CLINICAL MICROBIOLOGY. 2010;48(8):2729–33.
MLA
Mirabal, Niuris C, Sergio L Yzquierdo, Dihadenys Lemus, et al. “Evaluation of Colorimetric Methods Using Nicotinamide for Rapid Detection of Pyrazinamide Resistance in Mycobacterium Tuberculosis.” JOURNAL OF CLINICAL MICROBIOLOGY 48.8 (2010): 2729–2733. Print.
@article{1114298,
  abstract     = {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.},
  author       = {Mirabal, Niuris C and Yzquierdo, Sergio L and Lemus, Dihadenys and Madruga, Mariela and Milian, Yoslaine and Echemendia, Miguel and Takiff, Howard and Martin, Anandi and Van Der Stuyft, Patrick and Palomino, Juan Carlos and Montoro, Ernesto},
  issn         = {0095-1137},
  journal      = {JOURNAL OF CLINICAL MICROBIOLOGY},
  keyword      = {TUBERCLE-BACILLI,DRUG-RESISTANCE,ALTERNATIVE METHODS,RIFAMPICIN RESISTANCE,NITRATE REDUCTASE ASSAY,TESTING SUSCEPTIBILITY,PNCA MUTATIONS,PYRAZINOIC ACID,STRAINS,SYSTEM},
  language     = {eng},
  number       = {8},
  pages        = {2729--2733},
  title        = {Evaluation of colorimetric methods using nicotinamide for rapid detection of pyrazinamide resistance in Mycobacterium tuberculosis},
  url          = {http://dx.doi.org/10.1128/JCM.00311-10},
  volume       = {48},
  year         = {2010},
}

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