Real-time PCR quantification of bacterial adhesion to Caco-2 cells: Competition between bifidobacteria and enteropathogens
- 作者：Marco Candela ; Gerd Seibold ; Beatrice Vitali ; Sabrina Lachenmaier ; Bernhard J. Eikmanns and Patrizia Brigidi
- 关键词：Bifidobacterium ; Caco-2 cell adhesion ; Real-time PCR ; Enteropathogenic in vitro competition
- 刊名：Research in Microbiology
- 出版年：2005
- 期刊代码：75_09232508
- 类别：imm
- 出版时间：2005
- 卷：156
- 期：8
- 页码：887-895
- 文件大小：677 K
摘要
Probiotic bacteria play an important role in protecting the host from intestinal colonization of pathogenic bacteria. We have developed a new analytical approach based on a real-time PCR technique for quantifying Bifidobacterium adhesion to intestinal epithelial cells. Real-time PCR analysis showed that adhesion to enterocyte-like Caco-2 cells represented a variable phenotype in the genus Bifidobacterium, enabling classification of three adhesion behaviors: high adhesiveness (>40 bifidobacterial cells/Caco-2 cell); adhesiveness (5–40 bifidobacterial cells/Caco-2 cell); no adhesiveness (<5 bifidobacterial cells/Caco-2 cell). This molecular methodology was successfully used in competition studies in enteropathogens. All bifidobacterial strains examined evidenced displacement activity towards important enteropathogens (S. typhimurium, Y. enterocolitica and E. coli EPEC). Real-time PCR is a rapid, accurate and sensitive method for detecting and quantifying different bacterial genera and species simultaneously adhering to a epithelial cell monolayer.