The 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay as rapid colorimetric method for determination of antibiotic susceptibility of clinical Mycobacterium tuberculosis isolates in liquid medium.
We investigated the usefulness of a colorimetric method based on the reduction of a tetrazolium salt (XTT) for the susceptibility testing of clinical isolates of Mycobacterium tuberculosis to isoniazid, rifampin, rifabutin, ethambutol hydrochloride, ethionamide and streptomycin. The isolates and the ATCC reference strains reported as susceptible according to the agar dilution method approved by the National Committee for Clinical Laboratory Standards were found to be susceptible by the XTT colorimetric assay after times of incubation ranging between three days for rifampin and rifabutin to eight days for isoniazid. In comparison with other colorimetric methods reviewed in this article, the proposed assay is suitable for determining the susceptibility or resistance to most antituberculous drugs and, as a consequence of the water-solubility of the formazan yielded by reduction of XTT, additional steps such as the addition of extraction buffer and further incubation before the spectrophotometric analysis are not needed. The XTT reduction assay is an inexpensive, rapid and reliable screening method for the detection of susceptible, resistant and multidrug-resistant strains of M. tuberculosis and is an alternative to the costly performance of molecular or radiometric methods.
De Logu
Alessandro
A
Division of Medical Microbiology, Department of Biomedical Sciences and Technologies, University of Cagliari, Cagliari, Italy. adelogu@unica.it
Borgna
Rita
R
Uda
Patrizia
P
Sanna
Adriana
A
Pellerano
Maria Luisa
ML
Saddi
Barbara
B
D000995:Antitubercular Agents / Q000494:pharmacology*; D003124:Colorimetry / Q000379:methods; D024881:Drug Resistance, Bacterial; D006801:Humans; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D008826:Microbial Sensitivity Tests / Q000379:methods* / Q000592:standards; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000254:growth & development / Q000302:isolation & purification; D013778:Tetrazolium Salts; D018088:Tuberculosis, Multidrug-Resistant / Q000188:drug therapy / Q000382:microbiology
D000995:Antitubercular Agents; D007202:Indicators and Reagents; D013778:Tetrazolium Salts; C059087:2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino)carbonyl)-2H-tetrazolium hydroxide