Department of Analytical Chemistry, Nutrition and Food Science, School of Veterinary Sciences/College of Biotechnology, University of Santiago de Compostela, Lugo, Spain.
Electrophoresis. 2010 Mar;31(6):1116-27. doi: 10.1002/elps.200900591.
Histamine poisoning is caused by the consumption of fish and other foods that harbor bacteria possessing histidine decarboxylase activity. With the aim of preventing histamine formation, highly specific mass spectral fingerprints were obtained from the 16 major biogenic amine-producing enteric and marine bacteria by means of MALDI-TOF MS analysis. All bacterial strains analyzed exhibited specific spectral fingerprints that enabled its unambiguous differentiation. This technique also identified peaks common to certain bacterial groups. Thus, two protein peaks at m/z 4182+/-1 and 8363+/-6 were found to be present in all Enterobacteriaceae species analyzed except for Morganella morganii. Peaks at m/z 3635+/-1 and 7267+/-2 were specific to both M. morganii and Proteus spp. Biogenic amine-forming Proteus spp. exhibited three genus-specific peaks at m/z 3980, 7960+/-1 and 9584+/-2. The genus Photobacterium also showed three genus-specific peaks at m/z 2980+/-1, 4275+/-1 and 6578+/-1. The two histamine-producing Gram-positive bacteria Lactobacillus sp. 30A and Staphylococcus xylosus exhibited a few protein peaks in the 2000-7000 m/z range and could be easily distinguished from biogenic amine-forming Gram-negative bacteria. Clustering based on MALDI-TOF MS also exhibited a good correlation with phylogenetic analysis based on the 16S rRNA gene sequence, validating the ability of the MALDI-TOF technique to establish relationships between microbial strains and species. The approach described in this study leads the way toward the rapid and specific identification of major biogenic amine-forming bacteria based on molecular protein markers with a goal to the timely prevention of histamine food poisoning.
组胺中毒是由食用含有组氨酸脱羧酶活性的鱼类和其他食物引起的。为了防止组胺的形成，通过 MALDI-TOF MS 分析，从 16 种主要的产生物胺肠道和海洋细菌中获得了高度特异性的质谱指纹图谱。分析的所有细菌菌株都表现出特定的光谱指纹，能够明确地区分它们。该技术还鉴定了某些细菌群共有的峰。因此，在除摩根摩根菌外的所有肠杆菌科细菌中都发现了 m/z 4182+/-1 和 8363+/-6 两个蛋白质峰。m/z 3635+/-1 和 7267+/-2 两个峰特异性存在于摩根摩根菌和变形菌属细菌中。产生物胺的变形菌属细菌在 m/z 3980、7960+/-1 和 9584+/-2 处表现出三个属特异性峰。发光杆菌属也在 m/z 2980+/-1、4275+/-1 和 6578+/-1 处显示出三个属特异性峰。两种产组胺的革兰氏阳性菌乳杆菌 30A 和葡萄球菌 xylosus 在 2000-7000 m/z 范围内显示出一些蛋白质峰，很容易与产生物胺的革兰氏阴性菌区分开来。基于 MALDI-TOF MS 的聚类分析也与基于 16S rRNA 基因序列的系统发育分析有很好的相关性，验证了 MALDI-TOF 技术建立微生物菌株和物种之间关系的能力。本研究中描述的方法为基于分子蛋白标记物快速特异性鉴定主要产生物胺细菌开辟了道路，以期及时预防组胺食物中毒。
