Department of Analytical Chemistry, Nutrition and Food Science, School of Veterinary Sciences, University of Santiago de Compostela, Lugo, Spain.
Electrophoresis. 2013 Mar;34(6):877-87. doi: 10.1002/elps.201200532. Epub 2013 Feb 25.
The present study aims to compare two molecular technologies, 16S rRNA sequencing and MALDI-TOF MS, for bacterial species identification in seafood. With this aim, 70 reference strains from culture collections, including important seafood-borne pathogenic and spoilage bacterial species, and 50 strains isolated from commercial seafood products, were analysed by both techniques. Genomic analysis only identified the species of 50% of the isolated strains, proving to be particularly poor at identifying members of the Pseudomonas and Bacillus genera. In contrast, MALDI-TOF MS fingerprinting identified 76% of the strains at the species level. The mass spectral data were submitted to the SpectraBank database (http://www.spectrabank.org), making this information available to other researchers. Furthermore, cluster analysis of the peak mass lists was carried out with the web application SPECLUST and the calculated groupings were consistent with results determined by a phylogenetic approach that is based on the 16S rRNA sequences. However, the MALDI-TOF MS analysis demonstrated more discriminating potential that allowed for better classification, especially for the Pseudomonas and Bacillus genera. This is of importance with respect to the varying pathogenic and spoilage character at the intragenus and intraspecies level. In this sense, MALDI-TOF MS demonstrated to be a competent bacterial typing tool that extends phenotypic and genotypic approaches, allowing a more ample classification of bacterial strains.
本研究旨在比较两种分子技术,16S rRNA 测序和 MALDI-TOF MS,用于鉴定海鲜中的细菌种类。为此,本研究分析了 70 株来自培养物库的参考菌株,包括重要的海鲜源性致病菌和腐败菌,以及 50 株从商业海鲜产品中分离的菌株。基因组分析仅鉴定出 50%分离菌株的物种,证明其特别难以识别假单胞菌属和芽孢杆菌属的成员。相比之下,MALDI-TOF MS 指纹图谱可在物种水平上鉴定出 76%的菌株。质谱数据已提交给 SpectraBank 数据库(http://www.spectrabank.org),供其他研究人员使用。此外,使用 web 应用程序 SPECLUST 对峰质量列表进行了聚类分析,计算得出的分组与基于 16S rRNA 序列的系统发育方法确定的结果一致。然而,MALDI-TOF MS 分析表现出更好的区分潜力,允许更好的分类,特别是对于假单胞菌属和芽孢杆菌属。这对于不同的属内和种内的致病性和腐败性特征很重要。在这个意义上,MALDI-TOF MS 被证明是一种有能力的细菌分型工具,扩展了表型和基因型方法,允许更广泛的细菌菌株分类。