应用TGGE技术分析人肠道中双歧杆菌的组成

用温度梯度凝胶电泳(TGGE)技术结合16SrDNA克隆、测序,对健康人肠道中双歧杆菌的组成进行了分析。10例健康人肠道双歧杆菌的TGGE分析显示:人肠道内双歧杆菌的组成具有宿主特异性,不同人肠道双歧杆菌种的多样性和种类不同。通过对一健康儿童肠道双歧杆菌属特异性PCR扩增产物的测序及TGGE电泳行为的分析发现,该个体双歧杆菌TGGE图谱中的条带分别代表Bifidobacteriumbifidum、B.infantis、B.longum、B.adolescentis、B.pseudocatenulatum等种和一新种,其中B.pseudocatenulatum(假小链双歧杆菌)是大多数个体共有且较优势的种类。用传统培养方法只检出B.pseudocatenulatum和B.longum两种。基于双歧杆菌属特异性PCR基础上的TGGE方法结合16SrDNA克隆文库分析可较灵敏、直观地反映人肠道中双歧杆菌的组成。

Analysis of Bifidobacteria spp. Composition in human gut via temperature gradient gel electrophoresis (TGGE)

Temperature Gradient Gel Electrophoresis (TGGE) method combined with 16S rDNA clone library profiling was used to analyze Bifidobacteria spp. composition in human gut in this study. Bifidobacteria group-specific TGGE patterns of 10 healthy human individuals showed that the Bifidobacteria population in humans was host-specific. The genomic diversity and species composition of Bifidobacteria in different individual was dissimilar. Through sequencing and TGGE comigration analysis of the Bifidobacteria group-specific amplicons for one healthy boy, it was revealed that the TGGE bands of this individual represented species Bifidobacterium bifidum, B. infantis, B. longum, B. adolescentis, B. pseudocatenulatum and one potentially new species, respectively. B. pseudocatenulatum was the most common species of tested samples. While as control, only two species- B. pseudocatenulatum and B. longum were isolated using traditional culture method. Bifidobacteria group-specific PCR based TGGE method combined with 16S rDNA clone library profiling is a sensitive and effective approach to resolve the population structure of Bifidobacteria in microflora of human intestinal tract.