高黏附力双歧杆菌的筛选与鉴定

Screening and Identification of high adhesion Bifidobacteria to HT-29 Cell

利用体外细胞培养法,从实验室现有的10株双歧杆菌中筛选具有较强粘附能力的菌株。采用革兰氏染色法和平板计数法,评价了这10株双歧杆菌对HT-29细胞的粘附性能,通过测定这10株双歧杆菌的16S rRNA基因序列(16S rDNA)进行分类学鉴定。结果显示,WBBI01,WBBI02,WBIN03,WBBI06具有极强的黏附力,其黏附指数分别达1.97×103,2.17×103,3.57×103,1.88×103。经16S rDNA测序鉴定WBBI01,WBIN03,WBBI06均与两歧双歧杆菌S17有极高的同源性,而WBBI02属于长双歧杆菌。结果表明,双歧杆菌黏附性能具有明显的种属特异性,不同种属双歧杆菌的粘附能力相差极大,以两歧双歧杆菌的黏附性能最强,婴儿长双歧杆菌为最弱。

The adhesion capacity of ten Bifidobacteria strains in vitro was investigated. Gram staining and plate counting method were used to evaluate their adhesion capacity to HT-29 cell, and 16S rDNA sequencing analysis was carried out to identify the strain.. Based on the data in vitro, High attachment was observed in Bifidobacteria WBBI01, WBBI02, WBIN03, WBBI06, their adhesion index reached （bacterial ceUs/100 Caco-2 cells） 1.97x 103,2.17x10^3,3.57x10^3,1.88x10^3 repectively, while low attachment was shown in other strains. Identification of the strain indicates that the homology of Bifidobacteria WBBI01, WBIN03, WBBI06 with B. bifidum S17 were all above 99%, WBBI02 belongs to B. longum. The adhesion capacity varied considerably among different strains. B. bifidum WBIN03 was the most adhesive and the adhesion ability of Bifidobacteria WBAN07 was the weakest among all the tested strains.