Rapid detection of resistance to pyrazinamide in Mycobacterium tuberculosis using the resazurin microtitre assay

The resazurin microtitre plate assay (REMA) was evaluated to determine the susceptibility of Mycobacterium tuberculosis to pyrazinamide, and was compared with the broth microdilution method (BMM), the absolute concentration method (ACM) and pyrazinamidase (PZase) determination. Thirty-four M. tuberc...

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Bibliographic Details
Published in:	Journal of antimicrobial chemotherapy Vol. 66; no. 5; pp. 1044 - 1046
Main Authors:	ZANETTI CAMPANERUT, Paula Aline, DIAS GHIRALDI, Luciana, ESPINOZA SPOSITTO, Fernanda Luisa, NAKAMURA SATO, Daisy, FUJIMURA LEITE, Clarice Queico, HIROYUKI HIRATA, Mário, CRESPO HIRATA, Rosário Dominguez, FRESSATTI CARDOSO, Rosilene
Format:	Journal Article
Language:	English
Published:	Oxford Oxford University Press 01-05-2011 Oxford Publishing Limited (England)
Subjects:	Antibiotics. Antiinfectious agents. Antiparasitic agents Bacteria Bioassays Biological and medical sciences Comparative analysis Drug resistance Drug Resistance, Bacterial Humans Medical sciences Microbial Sensitivity Tests - economics Microbial Sensitivity Tests - methods Mycobacterium bovis Mycobacterium bovis - drug effects Mycobacterium bovis - metabolism Mycobacterium tuberculosis Mycobacterium tuberculosis - drug effects Mycobacterium tuberculosis - isolation & purification Mycobacterium tuberculosis - metabolism Oxazines - metabolism Pharmacology. Drug treatments Prescription drugs Pyrazinamide - pharmacology Sensitivity analysis Sensitivity and Specificity Time Factors Tuberculosis Tuberculosis - microbiology Xanthenes - metabolism susceptibility testing Drug susceptibility test Pyrazinamide mycobacteria TB Resistance Rapid technique Mycobacterium tuberculosis Mycobacteriales Mycobacteriaceae Bacteria Actinomycetes Antituberculous agent Antibacterial agent
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Summary:	The resazurin microtitre plate assay (REMA) was evaluated to determine the susceptibility of Mycobacterium tuberculosis to pyrazinamide, and was compared with the broth microdilution method (BMM), the absolute concentration method (ACM) and pyrazinamidase (PZase) determination. Thirty-four M. tuberculosis clinical isolates (26 susceptible and 8 resistant to pyrazinamide) and reference strains M. tuberculosis H37Rv ATCC 27294 and Mycobacterium bovis AN5 were tested. REMA and BMM showed 100% specificity and sensitivity when compared with ACM; BMM, however, demanded more reading time. The PZase determination assay showed 87.50% and 100% sensitivity and specificity, respectively. All tested methods in this preliminary study showed excellent sensitivity and specificity for the determination of pyrazinamide susceptibility of M. tuberculosis, but REMA was faster, low-cost and easy to perform and interpret. Additional studies evaluating REMA for differentiating pyrazinamide-resistant and -susceptible M. tuberculosis should be conducted on an extended panel of clinical isolates.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 ObjectType-Article-2 ObjectType-Feature-1
ISSN:	0305-7453 1460-2091
DOI:	10.1093/jac/dkr057