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J Med Microbiol 56 (2007), 733-737; DOI: 10.1099/jmm.0.46915-0
© 2007 Society for General Microbiology
ISSN 1473-5644

Use of a colorimetric assay to measure differences in cytotoxicity of Mycobacterium tuberculosis strains

Jorge Castro-Garza1, Hugo B. Barrios-García1,2, Delia Elva Cruz-Vega1, Salvador Said-Fernández1, Pilar Carranza-Rosales1, Carmen A. Molina-Torres3 and Lucio Vera-Cabrera3

1 División de Biología Celular y Molecular, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro Social, Monterrey, NL, Mexico

2 Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León, San Nicolás de los Garza, NL, Mexico

3 Servicio de Dermatología, Hospital Universitario ‘José E. González’, Monterrey, NL, Mexico

Correspondence
Jorge Castro-Garza
jorge.castro{at}biomedicas.net

Received 25 August 2006
Accepted 3 March 2007

Several techniques have been used to quantify the cytotoxicity produced by Mycobacterium tuberculosis bacilli on cell monolayers; however, they are semi-quantitative or time consuming. Herein, a method based on crystal violet (CV) uptake by THP-1 cell monolayers is described. This colorimetric method quantifies the cytotoxic effect as a function of the number of remaining cells after the infection with M. tuberculosis. Since this micro-organism is not stained by the dye, it does not produce a background that affects absorbance readings. As determined by CV assay (CVA), M. tuberculosis strain H37Rv destroyed 10.5 % of THP-1 cell monolayers at 24 h and 50.52 % at 72 h, while M. tuberculosis strains lacking the complete phospholipase C locus produced a reduced cytotoxic effect. The damage estimated by microscopy corresponded to the effect quantified by CVA. The results show that the use of CVA is a rapid, sensitive and reliable quantitative assay to measure the cytotoxicity of different M. tuberculosis strains.

Abbreviations: CV, crystal violet; CVA, crystal violet assay; PLC, phospholipase C; WST-1, 4-[3-4(iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate.






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