Total lipids were extracted from
Mycobacterium tuberculosis with chloroform-methanol (2:1), applied on a silica-gelel thin-layer plate, and developed with chloroform-methanol-acetone (90:10:5). Glycolipids were detected by spraying Anthrone-reagent and heating.Strain H37Rv of
M.tuberculosis showed four Anthrone-positive spots, namely trehalose-monomycolate, unidentified glycolipid, trehalose-dimycolate and GL-Rv, and strain H37Ra showed only two spots corresponding to trehalose-monomycolate and trehalose-dimycolate. (Fig.1) Other 4 laboratory-stock strains of
M.tuberculosis showed glycolipid-pattern identical with either of these two patterns.
One hundred and fifty-eight strains of
M.tuberculosis, isolated clinically from tuberculosis patients, were classified into 7 types according to their glycolipid-pattern. (Fig.2) Twenty-seven strains contained one more Anthrone-positive spot other than those of strain H37Rv. Pattern II was most frequently observed (60 strains), and then pattern I (33 strains), VI (29 strains), IV (13 strains), V (9 strains), VIE (8 strains), and III (6 strains). Pattern I corresponded to that of strain H37Ra and pattern VI corresponded to that of strain H37Rv.
Glycolipid-pattern did not correlate to clinical features of patients from whom the bacilli had been isolated.
A glycolipid, which moved to just under the solvent front, was a new glycolipid which has been found by us and designated as GL-Rv.Chemical structure of GL-Rv wasclarified by us as trehalose-polyacyl derivatives (no mycolic acid as the acyl residue) (Reference 9). Glycolipid-pattern was very stable and reproducible for each strain of
M. tuberculosis. The procedure is very simple and does not require any special instruments. Glycolipid-pattern of
M.tuberculosis may be very useful as a complementary method for the sub-typing of
M.tuberculosis.
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