S-Space College of Medicine/School of Medicine (의과대학/대학원) Laboratory Medicine (검사의학전공) Journal Papers (저널논문_검사의학전공)
Electrospray ionization-tandem mass spectrometry analysis of the mycolic acid profiles for the identification of common clinical isolates of mycobacterial species
- Song, Sang Hoon; Park, Kyoung Un; Lee, Jae Ho; Kim, Eui Chong; Song, Junghan; Kim, Jin Q.
- Issue Date
- ELSEVIER SCIENCE BV
- JOURNAL OF MICROBIOLOGICAL METHODS; Vol.77 2; 165-177
- Mycolic acid; Mass spectrometry; Electrospray ionization; Tuberculosis; Nontuberculosis mycobacteria
- Mycolic acids are unique and complex molecular structures found in mycobacterial species. In the present study, we investigated whether electrospray ionization-tandem mass spectrometry (ESI-MS/MS) can be used to identify mycobacterial species based on their mycolic acid profiles. Clinical isolates of Mycobacterium tuberculosis complex and 18 nontuberculosis mycobacterial (NTM) species identified by PCR-restriction fragment length polymorphism (RFLP) or real-time PCR were used for this analysis. Crude lipid extracts were prepared by saponifying 1-2 colonies of individual isolates of mycobacterial species and by chloroform and methanol (2:1, v/v) extraction. ESI-MS/MS in negative ion mode with high cone voltage and collision energy was used for mycolic acid profiling analysis. Combinatorial precursor ion scans of m/z 395, 367, and 339 in the range of m/z 1000-1400 resulted in spectra specific to individual mycobacteria. M. tuberculosis complex and M. pulveris showed major ions by performing precursor ion scans on m/z 395 and 367, while other NTM species showed major ions by performing scans on m/z 367 and 339. The different NTM species examined showed different species dependent mycolic acid profiles. In conclusion, we describe a rapid, reliable, and informative ESI-MS/MS protocol for mycolic acid profiling in mycobacterial species, which allows mycobacterial species to be easily identified in clinical laboratories. (C) 2009 Elsevier B.V. All rights reserved.
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