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Departments of Organic Chemistry and Microbiology, University of Newcastle, Newcastle upon Tyne NE1 7RU
ABSTRACT
Summary: introduction
Few simple and reliable tests are available for the differentiation of Mycobacterium, Nocardia and related taxa. There is some evidence to indicate that chemical markers will be useful for the identification of such actinomycetes (Goodfellow, 1973; Staneck & Roberts, 1974).
Lipids characteristic of nocardiae (LCN-As) can be detected by thin-layer chromatographic analysis of ethanol-diethyl ether (I: I, v/v) extracts of nocardiae, true corynebacteria and rhodochrous strains, but are generally absent from mycobacteria, rothiae and actinomadurae (Mordarska, Mordarski & Goodfellow, 1972; Goodfellow, 1973; Goodfellow et al. 1974). LCN-As have been identified as free mycolic acids (Goodfellow et al. 1973) with the general structural formula
where r+s+t+u lies between o and 4, and m, n, 0, p and q are all greater than I (Maurice, Vacheron & Michel, 1971; Minnikin, Pate1 & Goodfellow, 1974). The mycolic acids frommycobacteria also contain components with oxygen functions, e.g. >C=O, CH-OCH, COOH, in addition to the 3-hydroxyacid unit (Etkmadi, 1967). Thin-layer chromatography (t.1.c.) of the methyl esters of the total mycolic acids from mycobacteria can therefore be expected to produce a multiple spot pattern, while those from strains classified in the other taxa should only form a single spot.
This paper demonstrates that the pattern of spots corresponding to mycolic esters obtained on t.1.c. analysis of whole-organism methanolysates provides a means of distinguishing mycobacteria and other taxa containing mycolic acids. The technique is rapid, simple, requires no expensive equipment, and should be applicable in routine clinical laboratories.
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