Department of Milk Hygiene and Technology, Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences Brno, Palackého tř. 3a, CZ-61242 Brno, Czech Republic.
Int J Food Microbiol. 2012 Oct 1;159(2):107-14. doi: 10.1016/j.ijfoodmicro.2012.07.029. Epub 2012 Aug 10.
Lactobacilli are bacteria with important implications in food and feed fermentation. Detailed knowledge of the lactobacilli composition is of high relevance to food and health control, various industrial or biotechnological applications, etc., but accurate identification of the Lactobacillus species is not an easy task. In this study, three methods, i.e. polymerase chain reaction (PCR), amplified 16S rDNA restriction analysis with restrictase MseI (16S-ARDRA), and the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) were evaluated for their capabilities to identify Lactobacillus species. After comparison of the three methods on a set of 19 type/reference Lactobacillus strains, 148 strains isolated from dairy and meat products were characterized by PCR and MALDI-TOF MS. The strains were differentiated into nine and ten lactobacilli species by PCR and MALDI-TOF MS, respectively, with nine of these species being congruent. The success rates of species level assignment were 77% for PCR and 93% for MALDI-TOF MS that proved superior in the species identification. However, to differentiate between closely related Lactobacillus species, MALDI-TOF MS needs to be used in combination with genotypic techniques to achieve a more reliable identification.
乳杆菌是在食品和饲料发酵中具有重要意义的细菌。详细了解乳杆菌的组成对于食品和健康控制、各种工业或生物技术应用等具有高度相关性,但准确鉴定乳杆菌物种并非易事。在这项研究中,评估了聚合酶链反应(PCR)、限制性内切酶 MseI 扩增 16S rDNA 限制分析(16S-ARDRA)和基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)这三种方法识别乳杆菌物种的能力。在对一组 19 种类型/参考乳杆菌菌株进行比较后,用 PCR 和 MALDI-TOF MS 对从乳制品和肉类产品中分离出的 148 株菌株进行了特征描述。通过 PCR 和 MALDI-TOF MS 将这些菌株分为九种和十种乳杆菌,其中九种具有一致性。PCR 的种水平分配成功率为 77%,MALDI-TOF MS 的成功率为 93%,这表明 MALDI-TOF MS 在物种鉴定方面更具优势。然而,要区分密切相关的乳杆菌物种,需要将 MALDI-TOF MS 与基因技术结合使用,以实现更可靠的鉴定。