Kim Eiseul, Yang Seung-Min, Kim Hae-Yeong
Department of Food Science and Biotechnology, Institute of Life Sciences & Resources, Kyung Hee University, Yongin 17104, Korea.
Foods. 2021 Sep 7;10(9):2112. doi: 10.3390/foods10092112.
strains, isolated from raw milk and fermented dairy products, are closely related to the species that has beneficial probiotic properties. However, it is difficult to distinguish those using conventional methods. In this study, a unique gene was revealed to differentiate from other strains of the species and other species by pan-genome analysis, and a real-time PCR method was developed to rapidly and accurately detect the unique gene. The genome analysis of 141 genomes yielded an 17,978 pan-genome. Among them, 18 accessory genes were specifically present in five genomes of . The glycosyltransferase family 8 was identified as a unique gene present only in and not in 136 other genomes. A primer designed from the unique gene accurately distinguished in pure and mixed DNA and successfully constructed the criterion for the quantified standard curve in real-time PCR. The real-time PCR method was applied to 61 strains containing other species and distinguished with 100% accuracy. Also, the real-time PCR method was proven to be superior to the 16S rRNA gene method in the identification of .
从生牛奶和发酵乳制品中分离出的菌株与具有有益益生菌特性的物种密切相关。然而，使用传统方法很难区分这些菌株。在本研究中，通过泛基因组分析揭示了一个独特的基因，用于区分该物种的其他菌株和其他物种，并开发了一种实时PCR方法来快速准确地检测该独特基因。对141个基因组的分析产生了一个由17,978个基因组成的泛基因组。其中，18个辅助基因专门存在于该物种的5个基因组中。糖基转移酶家族8被鉴定为仅存在于该物种中而不存在于其他136个基因组中的独特基因。根据该独特基因设计的引物能够准确区分纯DNA和混合DNA中的该物种，并成功构建了实时PCR定量标准曲线的标准。实时PCR方法应用于包含该物种其他菌株的61个菌株，区分准确率达100%。此外，在该物种的鉴定中，实时PCR方法被证明优于16S rRNA基因方法。
