Microorganism identification by mass spectrometry and protein database searches
Microorganism identification by mass spectrometry and protein database searches
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DOI：
10.1021/ac990165u
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发表时间：
1999-07-15
影响因子：
7.4
通讯作者：
Fenselau, C
中科院分区：
文献类型：
--
作者：
Demirev, PA;Ho, YP;Fenselau, C
提出了一种快速鉴定微生物的方法，该方法利用了原核基因组和蛋白质序列数据库中包含的丰富信息。该方法基于通过完整或处理细胞的 MALDI TOF 质谱法确定一组离子的质量。随后通过搜索可通过互联网访问的蛋白质数据库来将集合中的每个离子与蛋白质以及蛋白质的有机来源相关联。对所有离子的列表进行卷积并对与匹配的离子相对应的生物体进行排序，从而识别出微生物。该方法已在枯草芽孢杆菌和大肠杆菌这两种基因组已完全测序的生物体上得到成功验证。该方法还通过微生物混合物的质谱、不同生长阶段的生物体的光谱以及来自其他实验室的光谱进行了鉴定测试。还讨论了 MALDI 基质制备、光谱再现性、污染物、质量范围和数据库搜索过程的测量精度等实验因素。与其他微生物鉴定 MS 方法相比，所提出的方法具有多个优点。
A method for rapid identification of microorganisms is presented, which exploits the wealth of information contained in prokaryotic genome and protein sequence databases. The method is based on determining the masses of a set of ions by MALDI TOF mass spectrometry of intact or treated cells. Subsequent correlation of each ion in the set to a protein, along with the organismic source of the protein, is performed by searching an Internet-accessible protein database. Convoluting the lists for all ions and ranking the organisms corresponding to matched ions results in the identification of the microorganism. The method has been successfully demonstrated on B, subtilis and E, coli, two organisms with completely sequenced genomes, The method has been also tested for identification from mass spectra of mixtures of microorganisms, from spectra of an organism at different growth stages, and from spectra originating at other laboratories. Experimental factors such as MALDI matrix preparation, spectral reproducibility, contaminants, mass range, and measurement accuracy on the database search procedure are addressed too. The proposed method has several advantages over other MS methods for microorganism identification.