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Journal of Clinical Microbiology, March 2008, p. 961-965, Vol. 46, No. 3
0095-1137/08/$08.00+0     doi:10.1128/JCM.01763-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Rapid Identification of Mycobacteria by Raman Spectroscopy

Department of Medical Microbiology and Infectious Diseases, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands,¹ Department of Medical Microbiology, Medical Centre Rijnmond-Zuid, Rotterdam, The Netherlands,² Center for Optical Diagnostics and Therapy, Department of Dermatology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands,³ Department of Medical Microbiology, Vlietland Hospital, Schiedam, The Netherlands,⁴ National Mycobacteria Reference Laboratory, National Institute for Public Health and the Environment, Bilthoven, The Netherlands⁵

Received 5 September 2007/ Returned for modification 20 November 2007/ Accepted 23 December 2007

A number of rapid identification methods have been developed to improve the accuracy for diagnosis of tuberculosis and to speed up the presumptive identification of Mycobacterium species. Most of these methods have been validated for a limited group of microorganisms only. Here, Raman spectroscopy was compared to 16S rRNA sequencing for the identification of Mycobacterium tuberculosis complex strains and the most frequently found strains of nontuberculous mycobacteria (NTM). A total of 63 strains, belonging to eight distinct species, were analyzed. The sensitivity of Raman spectroscopy for the identification of Mycobacterium species was 95.2%. All M. tuberculosis strains were correctly identified (7 of 7; 100%), as were 54 of 57 NTM strains (94%). The differentiation between M. tuberculosis and NTM was invariably correct for all strains. Moreover, the reproducibility of Raman spectroscopy was evaluated for killed mycobacteria (by heat and formalin) versus viable mycobacteria. The spectra of the heat-inactivated bacteria showed minimal differences compared to the spectra of viable mycobacteria. Therefore, the identification of mycobacteria appears possible without biosafety level 3 precautions. Raman spectroscopy provides a novel answer to the need for rapid species identification of cultured mycobacteria in a clinical diagnostic setting.

Published ahead of print on 3 January 2008.