| The balance of miciro-ecology in the intestinal flora of animals plays a major role in the health or the disease of body. It can make the balance be lost by ingestion antibiotics. However, Bifidobacteria can keep and restore the balance besides of probiotic functions——anti-tumor, immune increasing and anti-aging。Bifidobacterium BBMN01 strain, isolated from the faeces of old people aged one hundred years in Bama, Guangxi province, was investigated to high adhesive capability and endurance to stomach acid and bile acid. So it is necessary to detect its effect on mouse intestinal flora before it is used in the factory product.Presently, the detection methods of the effect of probiotics on the intestinal flora are traditional incubation, ordinary PCR and real-time PCR. In this study, a rapid and acute real-time quantitative PCR method with SYBR Green for detecting some kinds of intestinal bacteria by designing the primes of high specificity to amplily the aim gene. The reaction system is 30μL including SYBR(R) Green Realtime PCR Master Mix 15μL,prime 1μL,template 2μL,ddH2012μL. The reaction process is 95℃5min for denaturation,then 40 cycles of 95℃15s,59℃30s,72℃30s for bifidobacteria, 40 cycles of 95℃15s,61℃30s,72℃30s for lactobacilli and enteroccus, 35 cycles of 95℃15s,61℃30s,72℃30s for enterobaclli, and the standard curves are attained.Then,the effect of BBMN01 administration on the mouse intestinal flora were investigated by real-time PCR,the result showed that Bifidobacterium BBMN01 can keep the micro-ecologic balance of normal animals and restore the intestinal micro-ecologic banlance of animals, while the high dosage was the best and the bosom dosage was better than the lower,and that quantification results of the method and traditional incubation are consistent. In conclusion, real-time PCR is more sensitive,more specific and faster than traditional incubation method.
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