Youn So Youn, Ji Geun Eog, Han Yoo Ri, Park Myeong Soo
Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea.
Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University, Seoul 08826, Republic of Korea.
J Microbiol Biotechnol. 2017 May 28;27(5):909-915. doi: 10.4014/jmb.1610.10018.
BGN4 (BGN4) has many proven beneficial effects, including antiallergy and anticancer properties. It has been commercialized and used in several probiotic products, and thus strain-specific identification of this strain is very valuable for further strain-dependent physiological study. For this purpose, we developed novel multiplex polymerase chain reaction (PCR) primer sets for strain-specific detection of BGN4 in commercial products and fecal samples of animal models. The primer set was tested on seven strains of and 75 strains of the other species. The BGN4-specific regions were derived using megaBLAST against genome sequences of various databases and four sets of primers were designed. As a result, only BGN4 produced four PCR products simultaneously whereas the other strains did not. The PCR detection limit using BGN4-specific primer sets was 2.8 × 10 CFU/ml of BGN4. Those primer sets also detected and identified BGN4 in the probiotic products containing BNG4 and fecal samples from a BGN4-fed animal model with high specificity. Our results indicate that the PCR assay from this study is an efficient tool for the simple, rapid, and reliable identification of BGN4, for which probiotic strains are known.
BGN4具有许多已被证实的有益作用，包括抗过敏和抗癌特性。它已被商业化并用于多种益生菌产品中，因此对该菌株进行菌株特异性鉴定对于进一步开展依赖菌株的生理学研究非常有价值。为此，我们开发了新型多重聚合酶链反应（PCR）引物组，用于在商业产品和动物模型粪便样本中对BGN4进行菌株特异性检测。该引物组在7株[具体菌属未提及]和75株其他[具体菌属未提及]菌株上进行了测试。利用megaBLAST针对各种[具体菌属未提及]数据库的基因组序列推导BGN4特异性区域，并设计了四组引物。结果，只有BGN4同时产生了四种PCR产物，而其他菌株没有。使用BGN4特异性引物组的PCR检测限为2.8×10 CFU/ml的BGN4。这些引物组还能以高特异性检测和鉴定含有BNG4的益生菌产品以及来自喂食BGN4的动物模型的粪便样本中的BGN4。我们的结果表明，本研究中的PCR检测方法是一种用于简单快速且可靠地鉴定已知益生菌菌株BGN4的有效工具。
