Development of a firefly luciferase-based assay for determining antimicrobial susceptibility of Mycobacterium avium subsp. paratuberculosis

Paratuberculosis (Johne's disease) is a fatal disease of ruminants for which no effective treatment is available. Presently, no drugs against Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis), the causative agent of Johne's disease, are approved for use in livestock. Additi...

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Bibliographic Details
Published in:Journal of clinical microbiology 1999-02, Vol.37 (2), p.304-309
Main Authors: WILLIAMS, S. L, HARRIS, N. B, BARLETTA, R. G
Format: Article
Language:English
Subjects:
Anti-Bacterial Agents - pharmacology
Antibacterial agents
Antibiotics. Antiinfectious agents. Antiparasitic agents
Antitubercular Agents - pharmacology
Biological and medical sciences
Culture Media
Humans
Luciferases - genetics
Luciferases - metabolism
Luminescent Measurements
Medical sciences
Microbial Sensitivity Tests - methods
Mycobacteriology and Aerobic Actinomycetes
Mycobacterium avium paratuberculosis
Mycobacterium avium subsp. paratuberculosis - drug effects
Mycobacterium avium subsp. paratuberculosis - enzymology
Mycobacterium avium subsp. paratuberculosis - genetics
Pharmacology. Drug treatments
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Summary:Paratuberculosis (Johne's disease) is a fatal disease of ruminants for which no effective treatment is available. Presently, no drugs against Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis), the causative agent of Johne's disease, are approved for use in livestock. Additionally, M. paratuberculosis has been linked to a human chronic granulomatous ileitis (Crohn's disease). To assist in the evaluation of antimicrobial agents with potential activity against M. paratuberculosis, we have developed a firefly luciferase-based assay for the determination of drug susceptibilities. The microorganism used was M. paratuberculosis K-10(pYUB180), a clinical isolate carrying a plasmid with the firefly luciferase gene. The MICs determined by the broth macrodilution method were as follows: amikacin, 2 microg/ml; Bay y 3118, 0.015 microg/ml; clarithromycin, 1.25 microg/ml; D-cycloserine, 25 microg/ml; ethambutol, 20 microg/ml; and rifabutin, 0.5 microg/ml. The strain was resistant to isoniazid and kanamycin. The results obtained by the luciferase assay were identical or fell within 1 doubling dilution. These results suggest that a combination of amikacin, clarithromycin, and rifabutin may be the most efficacious therapy for the treatment of M. paratuberculosis infections and that the use of fluoroquinolone class of antibiotics deserves further consideration. We demonstrate that the luciferase drug susceptibility assay is reliable for M. paratuberculosis and gives results within 7 days, whereas the broth macrodilution method requires 14 days.
ISSN:0095-1137
1098-660X
DOI:10.1128/jcm.37.2.304-309.1999