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The use of the microplate alamar blue assay (MABA) to assess the susceptibility of Mycobacterium lepraemurium to anti-leprosy and other drugs

  • Original Article
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Journal of Infection and Chemotherapy

Abstract

Although murine leprosy is no longer a common illness, our understanding of the biology of this disease is incomplete. One particular example of this concerns the etiologic agent Mycobacterium lepraemurium (MLM). MLM is a fastidious microorganism that is difficult to grow in axenic media; in a way, this has hampered attempts to thoroughly study its physiological and metabolic characteristics. MLM is an obligate intracellular bacillus that invades macrophages and replicates profusely with a generation time that oscillates between 0.5 and 11 days. In the present study, we have successfully maintained MLM alive for more than 12 days in vitro, providing us with an opportunity to study its susceptibility to several anti-leprosy agents and other drugs. To achieve this, we used a fluorescence reduction assay of alamar blue (a resazurin) in a microplate format (microplate-alamar-blue-assay; MABA), which is a highly sensitive, practical, and inexpensive method for assaying cell viability. We found that MLM was highly susceptible to clofazimine and rifampicin and was less susceptible to streptomycin, thiacetazone, kanamycin, dapsone, and ethionamide, in that order. MLM was not susceptible to four plant triterpenoids (oleanolic acid, neolignan-c, sitosterol, and ursolic acid) for which bactericidal activity has been reported in M. tuberculosis. Because the MABA has high sensitivity, it can be used to monitor the activity of microorganisms that are difficult to cultivate (such as M. lepraemurium), in response to various drugs, thus offering a method to complement the study of murine leprosy, about which many questions remain unanswered.

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Acknowledgments

The authors thank Dr. Mónica Vargas-Lascari (ENCB/IPN) for providing the mice used in this study. Partial financial support came from Secretaría de Investigación y Posgrado (SIP 20100533 and SIP 20110401), IPN, México, Sistema Nacional de Investigadores (SNI, CONACYT, México), and Comisión de Operación y Fomento de las Actividades Académicas (COFAA, IPN, México).

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Authors and Affiliations

  1. Departamento de Inmunología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Carpio y Plan de Ayala, Colonia Santo Tomás, 11340, Mexico, D.F., Mexico

    Melby Mendoza-Aguilar, Laura Almaguer-Villagrán, Patricia Arce-Paredes, José Luis Cid-Gutiérrez & Oscar Rojas-Espinosa

  2. UIM Farmacología Productos Naturales Hospital Pediatría, CMN Siglo XXI, Mexico, Mexico

    Adelina Jiménez-Arellanes

Authors
  1. Melby Mendoza-Aguilar
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  2. Laura Almaguer-Villagrán
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  3. Adelina Jiménez-Arellanes
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  4. Patricia Arce-Paredes
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  5. José Luis Cid-Gutiérrez
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  6. Oscar Rojas-Espinosa
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Correspondence to Oscar Rojas-Espinosa.

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Mendoza-Aguilar, M., Almaguer-Villagrán, L., Jiménez-Arellanes, A. et al. The use of the microplate alamar blue assay (MABA) to assess the susceptibility of Mycobacterium lepraemurium to anti-leprosy and other drugs. J Infect Chemother 18, 652–661 (2012). https://doi.org/10.1007/s10156-012-0387-6

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  • DOI: https://doi.org/10.1007/s10156-012-0387-6

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