PCR法对益生菌产品中双歧杆菌的鉴别

Identification of Bifidobacterium in Probiotic Products by PCR

以含有婴儿双歧杆菌、两歧双歧杆菌和乳酸杆菌的复合型益生菌产品为目标,以PCR分子生物学鉴定方法为基础,利用Primer Premier 6软件进行引物设计,确定两歧双歧杆菌与婴儿双歧杆菌共属保守区域特异性引物,对DNA扩增产物进行琼脂糖凝胶电泳,并检查荧光条带位置,确认鉴别效果。结果表明,该方法可特异性鉴别产品中的婴儿双歧杆菌和两歧双歧杆菌,对近缘乳酸杆菌无扩增。同时,对复合型益生菌市售产品培养的双歧杆菌菌落进行PCR扩增并进行凝胶电泳,检查荧光条带位置均在128 bp左右,符合长度要求。本研究所建立的方法可以对复合型益生菌产品中双歧杆菌进行鉴别。

Targeting compound probiotic products contains Bifidobacterium infantis,Bifidobacterium bifitum and Lactobacillus.Basing on PCR molecular biology identification method,using Primer Premier 6 software to design the primers and confirming the conserve region-specific primers of bifidobacterium bifidum and bifidobacterium infantium.Checking the position of fluorescence bands for electrophoresis by AGAR gel of DNA amplification products to confirm the identification effect.The results showed that the method could specifically identify bifidobacterium bifidum and bifidobacterium infantium in the products,and there was no amplification for related Lactobacillus.After PCR amplification and AGAR gel electrophoresis,all cultured colonies of commercial product exhibited fluorescence bands at 128 bp position and conformed to the requirement.The method established in this study is able to identify bifidobacteria in mixed probiotics products.