Kaneko T, Kurihara H
Central Research Institute, Meiji Milk Products Co., Tokyo, Japan.
J Dairy Sci. 1997 Jul;80(7):1254-9. doi: 10.3168/jds.S0022-0302(97)76054-5.
The numbers of bifidobacteria in fecal samples were specifically determined by colony hybridization with the mixture of digoxigenin-labeled DNA probes that were prepared from whole chromosomal DNA of Bifidobacterium longum 6001 and Bifidobacterium adolescentis 6003. These DNA probes strongly hybridized with DNA of B. longum, B. adolescentis, Bifidobacterium breve, Bifidobacterium suis, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium angulatum, and Bifidobacterium animalis. Detectable positive signals with DNA of Bifidobacterium pseudolongum ssp. pseudolongum, Bifidobacterium catenulatum, and Bifidobacterium thermophilum were also found after hybridization. When dot-blot hybridization was performed with whole cells of 47 reference strains containing 11 species (16 strains) of bifidobacteria, all of the bifidobacteria tested could be specifically detected by using these DNA probes; Lactobacillus fermentum JCM 1173, however, showed a slight nonspecific signal. The counts of bifidobacteria by colony hybridization in the fecal samples of four of the five subjects were the same as the counts that were obtained by the conventional method using BL agar medium. Furthermore, no significant difference existed in the number of bifidobacteria that were determined by either method.
粪便样本中双歧杆菌的数量通过与地高辛标记的DNA探针混合物进行菌落杂交来特异性测定，这些探针由长双歧杆菌6001和青春双歧杆菌6003的全染色体DNA制备。这些DNA探针与长双歧杆菌、青春双歧杆菌、短双歧杆菌、猪双歧杆菌、婴儿双歧杆菌、两歧双歧杆菌、角双歧杆菌和动物双歧杆菌的DNA强烈杂交。杂交后还发现了与假长双歧杆菌亚种假长双歧杆菌、链状双歧杆菌和嗜热双歧杆菌的DNA可检测到的阳性信号。当用包含11种双歧杆菌（16株）的47株参考菌株的全细胞进行斑点杂交时，使用这些DNA探针可以特异性检测所有测试的双歧杆菌；然而，发酵乳杆菌JCM 1173显示出轻微的非特异性信号。五个受试者中有四个受试者粪便样本中通过菌落杂交测定的双歧杆菌数量与使用BL琼脂培养基的传统方法获得的数量相同。此外，两种方法测定的双歧杆菌数量没有显著差异。
