Carmen Collado Maria, Hernández Manuel
Dpto. Biotecnología. Area Microbiología, Universidad Politécnica de Valencia, Camino de Vera, 14, 46022 Valencia, Spain.
Microbiol Res. 2007;162(1):86-92. doi: 10.1016/j.micres.2006.09.007. Epub 2006 Nov 16.
The aim of this study was to identify and discriminate bacteria contained in commercial fermented milks with bifidobacteria by the use of amplified ribosomal DNA restriction analysis (ARDRA) and randomly amplified polymorphic DNA (RAPD) techniques. ARDRA of the 16S rDNA gene and RAPD were performed on 13 Lactobacillus strains, 13 Streptococcus and 13 Bifidobacterium strains isolated from commercial fermented milk. Lactobacillus delbrueckii, Streptococcus thermophilus and Bifidobacterium animalis isolates were identified by genus- and species-PCR and also, they were differentiated at genus and species level by ARDRA using MwoI restriction enzyme. The ARDRA technique allowed for the discrimination among these three related genus with the use of only one restriction enzyme, since distinctive profiles were obtained for each genus. Therefore it can be a simple, rapid and useful method for routine identification. Also, RAPD technique allowed the discrimination of all bacteria contained in dairy products, at genus- and strain-level by the performance of one PCR reaction.
本研究的目的是通过使用扩增核糖体DNA限制性分析（ARDRA）和随机扩增多态性DNA（RAPD）技术，鉴定和区分含有双歧杆菌的市售发酵乳中所含的细菌。对从市售发酵乳中分离出的13株乳酸杆菌、13株链球菌和13株双歧杆菌进行了16S rDNA基因的ARDRA和RAPD分析。通过属和种PCR鉴定了德氏乳杆菌、嗜热链球菌和动物双歧杆菌分离株，并且使用MwoI限制性内切酶通过ARDRA在属和种水平上对它们进行了区分。ARDRA技术仅使用一种限制性内切酶就能区分这三个相关属，因为每个属都获得了独特的图谱。因此，它可以成为一种简单、快速且有用的常规鉴定方法。此外，RAPD技术通过一次PCR反应就能在属和菌株水平上区分乳制品中所含的所有细菌。
