Real-time PCR quantification of bacterial adhesion to Caco-2 cells: competition between bifidobacteria and enteropathogens.
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.
Candela
Marco
M
Department of Pharmaceutical Sciences, CIRB-Center for Biotechnology, University of Bologna, Via Belmeloro 6, 40126 Bologna, Italy.
Seibold
Gerd
G
Vitali
Beatrice
B
Lachenmaier
Sabrina
S
Eikmanns
Bernhard J
BJ
Brigidi
Patrizia
P
D001422:Bacterial Adhesion / Q000502:physiology*; D001644:Bifidobacterium / Q000235:genetics / Q000502:physiology*; D001667:Binding, Competitive; D018938:Caco-2 Cells / Q000382:microbiology*; D002460:Cell Line; D004755:Enterobacteriaceae / Q000235:genetics / Q000502:physiology*; D004847:Epithelial Cells; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D013045:Species Specificity