Saito Yasuo, Sakamoto Mitsuo, Takizawa Satoru, Benno Yoshimi
Glico Dairy Products Co., Ltd., 2-14-1 Musashino, Akishima, Tokyo 196-0021, Japan.
FEMS Microbiol Lett. 2004 Feb 9;231(1):125-30. doi: 10.1016/S0378-1097(03)00951-0.
Real-time polymerase chain reaction (PCR) and nested reverse transcription (RT) PCR were applied to demonstrate the viability of lactobacilli in the feces of volunteers fed fermented milk containing lactobacilli. Two sets of specific primers and a TaqMan probe for real-time PCR were constructed using the S-layer gene as a target. After fermented milk ingestion, Lactobacillus helveticus GCL1001 was detected in the feces of 12 volunteers over a few days, with the maximum number being between 10(4.5) and 10(7.8) cells g(-1) of feces. Moreover, mRNA from this strain was detected in the feces of all volunteers by nested RT-PCR. The results show that these methods are applicable to the demonstration of bacterial viability in feces, and that ingested L. helveticus GCL1001 can survive through the gastrointestinal tract.
应用实时聚合酶链反应（PCR）和巢式逆转录（RT）PCR来证明食用含乳酸菌发酵乳的志愿者粪便中乳酸菌的生存能力。使用表层蛋白基因作为靶点构建了两组用于实时PCR的特异性引物和一个TaqMan探针。食用发酵乳后，在几天内于12名志愿者的粪便中检测到瑞士乳杆菌GCL1001，粪便中该菌的最大数量在每克粪便10(4.5)至10(7.8)个细胞之间。此外，通过巢式RT-PCR在所有志愿者的粪便中检测到了该菌株的mRNA。结果表明，这些方法适用于证明粪便中细菌的生存能力，并且摄入的瑞士乳杆菌GCL1001能够在胃肠道中存活。
