A real-time quantitative PCR assay for evaluating Clostridium difficile adherence to differentiated intestinal Caco-2 cells.
Herein we describe a real-time quantitative PCR assay for evaluating the adherence of Clostridium difficile to differentiated human intestinal Caco-2 cells. Our investigations demonstrated that the method, employing the C. difficile-specific triose-phosphate isomerase gene, is as reliable but less time-consuming than counting c.f.u. We conclude that the method will be useful for evaluating the role of host cell adherence in the pathogenesis of C. difficile infection.
Dingle
Tanis
T
Department of Microbiology & Infectious Diseases, Faculty of Medicine, University of Calgary, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada.
Mulvey
George L
GL
Department of Microbiology & Infectious Diseases, Faculty of Medicine, University of Calgary, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada.
Humphries
Romney M
RM
Department of Microbiology & Infectious Diseases, Faculty of Medicine, University of Calgary, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada.
Armstrong
Glen D
GD
Department of Microbiology & Infectious Diseases, Faculty of Medicine, University of Calgary, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada.
D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics; D018938:Caco-2 Cells; D016360:Clostridioides difficile / Q000302:isolation & purification / Q000472:pathogenicity*; D015169:Colony Count, Microbial / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics; D004847:Epithelial Cells / Q000382:microbiology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D014305:Triose-Phosphate Isomerase / Q000235:genetics
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D014305:Triose-Phosphate Isomerase