MALDITOF质谱技术分析与鉴定病原细菌研究.docVIP

微生物学通报Microbiology HYPERLINK mailto:tongbao@ tongbao@MAR 20, 2009, 36(3): 416 微生物学通报 Microbiology HYPERLINK mailto:tongbao@ tongbao@ MAR 20, 2009, 36(3): 416~426 ? 2009 by Institute of Microbiology, CAS 研究快报 MALDI-TOF 质谱技术分析与鉴定病原细菌研究 其布勒哈斯 1,2 田世民 2* 邹明强 2 贺银凤 1 (1. 内蒙古农业大学 内蒙古 呼和浩特 艳 2 赵贵明 2 陈 010018) (2. 中国检验检疫科学研究院 北京 100123) 摘 要: 基于基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术分析供试细菌并获得其 蛋白质 图 谱 , 确定了与 质 谱 分析有关的最 佳 培 养 条 件 和 样 品 前 处理方法 , 建立 了 应 用 MALDI-TOF-MS 鉴定细菌的方法。通过对供试的不同细菌属、种和亚种的 12 株植物病原细菌全 细胞分析后的质谱图结果表明, 根据特异性或不同的质量离子峰, MALDI-TOF-MS 能进行在细菌 属、种、亚种和菌株水平上的区分和鉴定。该方法准确、灵敏度高, 且分析过程简单快速, 具有 广泛的开发和应用意义。 关键词: MALDI-TOF MS, 细菌, 质量指纹图谱, 鉴定 Analysis and Identification of Pathogenic Bacteria by MALDI-TOF Mass Spectrometry Qibulehasi1,2 TIAN Shi-Min2* ZOU Ming-Qiang2 HE Yin-Feng1 CHEN Yan2 ZHAO Gui-Ming2 (1. Inner Mongolian Agricultural University, Hohhot, Inner Mongolia 010018, China) (2. Chinese Academy of Inspection and Quarantine, Beijing 100123, China) Abstract: Based on Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), protein mass spectrometric profiles for bacteria were obtained, method for analysis of the bacteria was developed, and culture conditions and pre-treatments for bacteria samples were optimized. Results from analysis of twelve plant pathogenic bacteria of different genus, species and subspecies, and comparison of the protein mass spectrometric profiles showed that, MALDI-TOF-MS was able to identify the plant bacteria at the different levels of bacterial strains. This analytical method has the advantages of simple process, high sensitivity and accuracy, showing the useful significances. Keywords: MALDI-TOF-MS, Bacteria, Mass finger-printing, Identification 基质 辅助激 光解析 电离 飞行时 间质 谱 (Matrix- assisted laser desorption/ionization time of flight mass spectrometry, MALDI-TOF-MS)是近些年发展起来 的用于微生 物(细菌、酵母菌和真菌等 )快速区分和 鉴定的一项新技术, 其基本原理为：将微生物样品 与等量的基质溶液混合或分别点加在样品板上, 溶 剂挥发后形成样品与基质的共结晶; 利用激光作为 能量来源辐射结晶体, 基质从激光中吸收能