Bioluminescent Mycobacterium aurum expressing firefly luciferase for rapidand high throughput screening of antimycobacterial drugs in vitro and in infected macrophages
Dk. Deb et al., Bioluminescent Mycobacterium aurum expressing firefly luciferase for rapidand high throughput screening of antimycobacterial drugs in vitro and in infected macrophages, BIOC BIOP R, 279(2), 2000, pp. 457-461
Citations number
14
Categorie Soggetti
Biochemistry & Biophysics
Journal title
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
The slow growth and highly infectious nature of Mycobacterium tuberculosis
is a limiting factor in its use as test organism in high throughput screeni
ng for inhibitory compounds. To overcome these problems, use of surrogate s
trains and reporter genes have been considered. In this study, we have inve
stigated the application of a fast growing nonpathogenic M. aurum expressin
g firefly luciferase in rapid screening of antituberculosis compounds in vi
tro and in infected macrophages using bioluminescence assay. The assay is b
ased on luminescence determination using luciferin as substrate. Inhibition
of bioluminescence was obtained with frontline antimycobacterial drugs lik
e streptomycin, rifampicin, isoniazid, ethambutol, ofloxacin, and sparfloxa
cin at their reported MICs. Inhibition could be observed as early as 2 h in
vitro and within 24 h in infected macrophages. The system can reliably be
used in high throughput screening. (C) 2000 Academic Press.