电喷雾电离
霉酸
串联质谱法
化学
电喷雾
质谱法
分枝杆菌
生物
色谱法
细菌
遗传学
作者
Sang Hoon Song,Kyoung Un Park,Jae Ho Lee,Eui Chong Kim,Jin Q Kim,Jae-Joon Song
标识
DOI:10.1016/j.mimet.2009.01.023
摘要
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.
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