Comparison of two rapid colorimetric methods for determining resistance of mycobacterium tuberculosis to rifampin, isoniazid, and streptomycin in liquid medium.
The usefulness of two colorimetric methods for the determination of the susceptibility or resistance of Mycobacterium tuberculosis to rifampin, streptomycin, and isoniazid in liquid medium based on the reduction of 2,3-bis(2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide (XTT) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was investigated. The agar proportion method was used as the reference method. Results obtained indicate that the sensitivity of the XTT reduction assay for the detection of rifampin resistance was comparable to that observed, and previously described, for the MTT assay. However, the reduction of XTT yields a water-soluble formazan that can be easily quantified without performing additional steps such as addition of lysing buffer and solubilization. Furthermore, the colorimetric assays, based on the reduction of XTT and MTT for the detection of isoniazid and streptomycin resistance in Mycobacterium tuberculosis, were standardized. The inhibition of MTT and
De Logu
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A
Dipartimento di Scienze Chirurgiche e Trapianti d'Organo, Università di Cagliari, Italy. adelogu@vaxca1.unica.it
Uda
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Pellerano
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ML
Pusceddu
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MC
Saddi
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B
Schivo
M L
ML
D000904:Antibiotics, Antitubercular / Q000494:pharmacology; D000995:Antitubercular Agents / Q000494:pharmacology*; D003124:Colorimetry / Q000379:methods*; D004396:Coloring Agents / Q000378:metabolism; D004352:Drug Resistance, Microbial; D007538:Isoniazid / Q000494:pharmacology; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000378:metabolism; D010084:Oxidation-Reduction; D012293:Rifampin / Q000494:pharmacology; D012680:Sensitivity and Specificity; D013307:Streptomycin / Q000494:pharmacology; D013997:Time Factors; D018088:Tuberculosis, Multidrug-Resistant
D000904:Antibiotics, Antitubercular; D000995:Antitubercular Agents; D004396:Coloring Agents; D007538:Isoniazid; D012293:Rifampin; D013307:Streptomycin