Quantitative PCR with 16S rRNA-Gene-targeted species-specific primers for analysis of human intestinal bifidobacteria
Quantitative PCR with 16S rRNA-Gene-targeted species-specific primers for analysis of human intestinal bifidobacteria
复制标题
DOI：
10.1128/aem.70.1.167-173.2004
复制
发表时间：
2004-01-01
影响因子：
4.4
通讯作者：
Tanaka, R
中科院分区：
文献类型：
--
作者：
Matsuki, T;Watanabe, K;Tanaka, R
将双歧杆菌属种特异性引物与实时荧光定量PCR技术相结合，建立了一种高灵敏度的双歧杆菌定量PCR检测方法，并应用于人体肠道双歧杆菌的分布分析。从培养的双歧杆菌中提取的连续稀释的DNA的实时PCR检测对于每个PCR测定范围为10（6）至10个细胞的细胞计数是线性的。还发现，当粪便中双歧杆菌的浓度为>10（6）个细胞/g粪便时，该方法适用于检测粪便中的双歧杆菌。关于肠道植物群中双歧杆菌的种类分布，通过检查从46名健康成人的粪便中提取的DNA，发现双歧杆菌属的双歧杆菌群、链状双歧杆菌群和长双歧杆菌是三种优势种。我们还研究了在8个月的时间内，6名健康成年人的肠道植物群中双歧杆菌的种群和组成的变化。结果表明，在整个试验期间，双歧杆菌植物群的组成基本稳定。
A highly sensitive quantitative PCR detection method has been developed and applied to the distribution analysis of human intestinal bifidobacteria by combining real-time PCR with Bifidobacterium genus- and species-specific primers. Real-time PCR detection of serially diluted DNA extracted from cultured bifidobacteria was linear for cell counts ranging from 10(6) to 10 cells per PCR assay. It was also found that the method was applicable to the detection of Bifidobacterium in feces when it was present at concentrations of >10(6) cells per g of feces. Concerning the distribution of Bifidobacterium species in intestinal flora, the Bifidobacterium adolescentis group, the Bifidobacterium catenulatum group, and Bifidobacterium longum were found to be the three predominant species by examination of DNA extracted from the feces of 46 healthy adults. We also examined changes in the population and composition of Bifidobacterium species in human intestinal flora of six healthy adults over an 8-month period. The results showed that the composition of bificlobacterial flora was basically stable throughout the test period.