Krishnamurthy T, Ross P L, Rajamani U
U.S. Army Edgewood RD&E Center, APG, MD 21010-5423, USA.
Rapid Commun Mass Spectrom. 1996;10(8):883-8. doi: 10.1002/(SICI)1097-0231(19960610)10:8<883::AID-RCM594>3.0.CO;2-V.
The proteins isolated from the whole cells of bacterial pathogens and related non-pathogenic simulants were analyzed directly, with minimal sample preparation by matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometry. Inspection of mass spectrometric profiles obtained from direct MALDI-MS analysis of the protein extracts revealed specific biomarkers for individual bacterial cells. The observed biomarkers enabled us not only to detect pathogenic bacteria (Bacillus anthracis, Yersinia pestis and Brucella meliteusis), but also to distinguish them from the corresponding non-pathogenic species. By examining a series of strains of several Bacillus species (anthracis, thuringiensis, cereus and subtilis), it was possible to derive genus, species and strain-specific biomarkers from the measured molecular masses of the intact proteins. Additional series of biomarkers were obtained from direct mass spectrometric analysis of tryptic digests of the protein extracts. The application of this technique for rapid chemotaxonomic classification of microorganisms is demonstrated.
对从细菌病原体全细胞及相关非致病性模拟物中分离出的蛋白质进行直接分析，通过基质辅助激光解吸/电离（MALDI）飞行时间质谱法进行最少的样品制备。对蛋白质提取物直接进行MALDI-MS分析所获得的质谱图检查揭示了单个细菌细胞的特异性生物标志物。观察到的生物标志物使我们不仅能够检测致病细菌（炭疽芽孢杆菌、鼠疫耶尔森菌和马尔他布鲁氏菌），还能将它们与相应的非致病物种区分开来。通过检查几种芽孢杆菌属物种（炭疽芽孢杆菌、苏云金芽孢杆菌、蜡样芽孢杆菌和枯草芽孢杆菌）的一系列菌株，可以从完整蛋白质的测量分子量中得出属、种和菌株特异性生物标志物。从蛋白质提取物的胰蛋白酶消化物的直接质谱分析中获得了更多系列的生物标志物。展示了该技术在微生物快速化学分类学分类中的应用。
