35S-Methioninei摂取後の薄層クロマトグラフィーによる抗酸菌菌種同定法に関する知見補遺

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Tsukamura and Mizuno have reported that thin-layer chromatography of ethyl ether- ethanol-soluble fraction of mycobacteria after incubation with 35S-methionine is useful for differentiation among mycobacterial species, and that distribution of radioactive spots in thin- layer chromatograms show a pattern specific for the species, although it has been observed that M. gordonae shows various patterns. In the present, study, the results on the mycobacteria previously not studied are reported. The majority of the mycobacteria studied in the present study belong to rapidly growing, scotochromogenic mycobacteria. The strains used are shown in Table 1. The methods used were the same as reported previously. The results obtained are shown in Figs. 1 to 10 and in Table 2. The following rapidly growing, scotochromogenic mycobacteria were shown to present an almost species-specific pattern of the distribution of radioactive spots in thin-layer chromato grams: M. chubuense; M. aichiense; M. obuense; M. rhodesiae; M. gilvurn; M. duvalii; M. phlei; M. parafortuitum; M. aurum; M. neoaurum. Only a few exceptions were observed. A strain of M. neoaurum lacked the spot f, although other strains of this species showed the spot (Fig. 3, F). A strain of M. aurum showed the same pattern as of M. parafortuitum (Fig. 4, F). M. parafortuitum and M. phlei showed the same pattern (Fig. 3 and 4). In contrast to the above species, M. flavescens showed different patterns within the species (Fig.5). However, the patterns could be subgrouped to approximately two groups: the first type (Fig.5, upper) contained the type strain of M. flavescens, and the second (Fig.5, lower) contained the type strain of the species originally received as M. gallinarum. Among slowly growing mycobacteria tested in the study, M. kansasii, M. marinum and M. nonchromogenicum were shown to be a homogeneous species so far viewed from the pattern, showing the same pattern within the strains of each species. M. triviale could be differentiated from M. nonchromogenicum, showing a different pattern, and M. terrae was differentiated from M. nonchromogenicum, as the former lacked the spot f. The pattern of M. malmoense resembled to that of M. avium-intracellulare complex. The strains of M. szulgai showed various patterns and seemed to be heterogenous when viewed from the pattern. M. kansasii and M. marinum could be differentiated clearly by a difference of the pattern. The former showed the spot f and the latter lacked this spot. To test the nature of the spot f, the concentrates of the ethyl ether-ethanol-soluble frac tion of the strains of these species were added with petroleum ether and a little amount of water, and a separated petroleum ether-layer was concentrated under reduced pressure and subjected to the thin-layer chromatography in the same manner. The petroleum ether-extracts of M. kansasii showed only the spot f, and those of M. marinum showed no spot (Fig. 11). The results show that a substance showing the spot f is soluble in petroleum ether, whereas other substances showing the other spots are not soluble in petroleum ether. The test of the distribution of radioactive spots in thin-layer chromatography of the ethyl ether-ethanol-soluble fraction of mycobacteria has been shown to be useful for the differentiation among mycobacterial species. 1) Tsukamura, M. and Mizuno, S.: Int. J. Syst. Bacteriol., 25: 271, 1975. 2) Tsukamura, M. and Mizuno, S.: Kekkaku, 53: 85, 1978.
日本結核病学会の論文

35S-Methioninei摂取後の薄層クロマトグラフィーによる抗酸菌菌種同定法に関する知見補遺

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