Abstract
In order to differentiate between mycobacterial species, thin-layer chromatography of petroleum ether-soluble lipids was carried out, using solvent system consisting of petroleum ether: diethyl ether: acetic acid (90:10:1, involume). Wet weight of 100 mg of test organism was extracted twice with 2.0 ml of petroleum ether, and the extracts were combined and dried. The dried material was dissolved in 0.1 ml of petroleum ether and placed on a thin-layer of Silica Gel H (Merck, Darmstadt, Germany; 20 by 20 cm, 0.25 mm thick). After development, the lipid spots were visualized by spraying a 5% phosphorus molybdatemethanol solution and by heating them at 90°C for 10 minutes. The lipids appeared as brown or green spots.
The following slowly-growing-mycobacteria were differentiated from each other, showing a pattern specific for the species: Mycobacterium bovis; M, kansasii; M. marinum; M. avium; M. scrofulaceum. In contrast, M. nonchromogenicum, M. gordonae and M. intracellular (3 strains) showed the same pattern and were not differentiated from each other by this method.
The following rapidly-growing-mycobacteria were differentiated from each other, showing a pattern specific for the species: Mycobacterium fortuitum; M, chelonei subsp. chelonei; M. chitae; M, smegmatis; M. parafortuitum; M, thermoresistibile. In contrast, M. phlei and M. flavescens, and M. vaccae and M. aurum showed similar patterns respectively. Three species of the genus Rhodococcus showed the same pattern as shown by M. vaccae and M. aurum.
