Mättö J, Malinen E, Suihko M-L, Alander M, Palva A, Saarela M
VTT Biotechnology, Espoo, Finland.
J Appl Microbiol. 2004;97(3):459-70. doi: 10.1111/j.1365-2672.2004.02340.x.
The aim of the present study was to compare several molecular methods for the identification and genotyping of bifidobacteria, and further to investigate genetic heterogeneity and functional properties of bifidobacterial isolates from intestinal samples of Finnish adult subjects.
A total of 153 intestinal bifidobacterial isolates were included in initial screening and 34 isolates were further characterized. Identification results obtained with PCR-ELISA and ribotyping were well in accordance with each other, while randomly amplified polymorphic DNA (RAPD) gave tentative identification only to Bifidobacterium bifidum and to 65% of the B. longum isolates. The most commonly detected species were B. longum biotype longum followed by B. adolescentis and B. bifidum. In addition, B. animalis (lactis), B. angulatum and B. pseudocatenulatum were found. Ribotyping and pulsed-field gel electrophoresis (PFGE) proved to be discriminatory methods for bifidobacteria, but also RAPD revealed intraspecies heterogeneity. Besides two B. animalis (lactis) isolates with very close similarity to a commercially available probiotic strain, none of the intestinal isolates showed optimal survival in all tolerance (acid, bile and oxygen) or growth performance tests.
Several species/strains of bifidobacteria simultaneously colonize the gastrointestinal tract of healthy Finnish adults and intestinal Bifidobacterium isolates were genetically heterogeneous. Functional properties of bifidobacteria were strain-dependent.
Applicability of ribotyping with the automated RiboPrinter System for identification and genotyping of bifidobacteria was shown in the present study.
本研究旨在比较几种用于双歧杆菌鉴定和基因分型的分子方法,并进一步研究来自芬兰成年受试者肠道样本的双歧杆菌分离株的遗传异质性和功能特性。
共有153株肠道双歧杆菌分离株纳入初步筛选,34株分离株进一步进行特性分析。PCR-ELISA和核糖体分型获得的鉴定结果相互吻合,而随机扩增多态性DNA(RAPD)仅对两歧双歧杆菌和65%的长双歧杆菌分离株给出初步鉴定。最常检测到的菌种是长双歧杆菌长亚种,其次是青春双歧杆菌和两歧双歧杆菌。此外,还发现了动物双歧杆菌(乳亚种)、角双歧杆菌和假链状双歧杆菌。核糖体分型和脉冲场凝胶电泳(PFGE)被证明是双歧杆菌的鉴别方法,但RAPD也揭示了种内异质性。除了两株与市售益生菌菌株非常相似的动物双歧杆菌(乳亚种)分离株外,没有一株肠道分离株在所有耐受性(酸、胆汁和氧气)或生长性能测试中表现出最佳存活能力。
几种双歧杆菌菌种/菌株同时定殖于健康芬兰成年人的胃肠道,肠道双歧杆菌分离株具有遗传异质性。双歧杆菌的功能特性具有菌株依赖性。
本研究表明了使用自动核糖体分型仪系统进行核糖体分型在双歧杆菌鉴定和基因分型中的适用性。