Sato Jun, Nakayama Motokazu, Tomita Ayumi, Sonoda Takumi, Hasumi Motomitsu, Miyamoto Takahisa
Safety Science Research, R&D, Kao Corporation, Ichikai, Tochigi, Japan.
Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Hakozaki, Higashi-ku, Fukuoka, Japan.
PLoS One. 2017 Oct 11;12(10):e0186327. doi: 10.1371/journal.pone.0186327. eCollection 2017.
In order to establish rapid and accurate typing method for Bacillus coagulans strains which is important for controlling in some canned foods and tea-based beverages manufacturing because of the high-heat resistance of the spores and high tolerance of the vegetative cells to catechins and chemicals, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and repetitive-PCR (rep-PCR) were evaluated. For this purpose, 28 strains of B. coagulans obtained from various culture collections were tested. DNA sequence analyses of the genes encoding 16S rRNA and DNA gyrase classified the test strains into two and three groups, respectively, regardless of their phenotypes. Both MALDI-TOF MS and rep-PCR methods classified the test strains in great detail. Strains classified in each group showed similar phenotypes, such as carbohydrate utilization determined using API 50CH. In particular, the respective two pairs of strains which showed the same metabolic characteristic were classified into the same group by both MALDI-TOF MS and rep-PCR methods separating from the other strains. On the other hand, the other strains which have the different profiles of carbohydrate utilization were separated into different groups by these methods. These results suggested that the combination of MALDI-TOF MS and rep-PCR analyses was advantageous for the rapid and detailed typing of bacterial strains in respect to both phenotype and genotype.
凝结芽孢杆菌菌株的芽孢具有高耐热性，其营养细胞对儿茶素和化学物质具有高耐受性，这对于某些罐头食品和茶饮料生产中的控制至关重要。为了建立快速准确的凝结芽孢杆菌菌株分型方法，对基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）和重复PCR（rep-PCR）进行了评估。为此，测试了从各种培养物保藏中心获得的28株凝结芽孢杆菌。编码16S rRNA和DNA促旋酶的基因的DNA序列分析分别将测试菌株分为两组和三组，与它们的表型无关。MALDI-TOF MS和rep-PCR方法都对测试菌株进行了非常详细的分类。每组中分类的菌株表现出相似的表型，例如使用API 50CH测定的碳水化合物利用情况。特别是，具有相同代谢特征的两对菌株通过MALDI-TOF MS和rep-PCR方法被分类到同一组中，与其他菌株分开。另一方面，具有不同碳水化合物利用谱的其他菌株通过这些方法被分为不同的组。这些结果表明，MALDI-TOF MS和rep-PCR分析的组合对于细菌菌株在表型和基因型方面的快速和详细分型是有利的。
