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Journal of Clinical Microbiology, September 1998, p. 2477-2480, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Improved Identification of Mycobacteria by Using the Microbial Identification System in Combination with Additional Trimethylsulfonium Hydroxide Pyrolysis

K.-Dieter Müller,1,* Ernst N. Schmid,1 and Reiner M. Kroppenstedt2

Institut für Medizinische Mikrobiologie, Universität GH-Essen, D-45122 Essen,1 and German Collection of Microorganisms and Cell Cultures, D-38124 Braunschweig,2 Germany

Received 21 January 1998/Returned for modification 9 March 1998/Accepted 16 June 1998

The MIDI automated Microbial Identification System (MIS) uses gas chromatography (GC) analysis of whole-cell fatty acid methyl esters (FAMEs) between 9 and 20 carbons in length to characterize a wide range of bacterial genera and species, including mycobacteria. Mycolic acid cleavage products (MACPs) with chain lengths of C22 to C26 are not released by MIDI sample preparation of mycobacteria. Therefore, the MIS library search report often matches several mycobacterial species without any significant difference in the similarity indices. The problem is solved by adding trimethylsulfonium hydroxide (TMSH) instead of sodium sulfate in the last step of sample preparation, thus allowing the identification of MACPs in addition to FAMEs. Only one GC run parameter has to be changed: the temperature program must be extended from 260 to 310°C. The MIS library search report for the identification of bacteria is not disturbed by TMSH. The combination of conventional library search report with the information of typical MACP patterns yields significantly better discrimination of mycobacterial species than the MIDI method allows.

* Corresponding author. Mailing address: Institut für Med. Mikrobiologie, Universität -GH Essen, Hufelandstr. 55, D-45122 Essen, Germany. Phone: 49-201-723-3520. Fax: 49-201-723-5729. E-mail: karl-dieter.mueller{at}uni-essen.de.

Journal of Clinical Microbiology, September 1998, p. 2477-2480, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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