Quantification of specific E. coli in gut mucosa from Crohn's disease patients

• Published in: Journal of microbiological methods Vol. 86; no. 1; pp. 111 - 114
• Main Authors: Jensen, Stina Rikke, Fink, Lisbeth Nielsen, Struve, Carsten, Sternberg, Claus, Andersen, Jens Bo, Brynskov, Jørn, Nielsen, Ole Haagen, Brix, Susanne
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.07.2011; Elsevier
• Subjects: Adherent-invasive E. coli; Bacterial quantification; Biological and medical sciences; Crohn disease; Crohn Disease - microbiology; Crohn's disease; digestive tract mucosa; DNA Primers - genetics; Escherichia coli; Escherichia coli - genetics; Escherichia coli - isolation & purification; Fundamental and applied biological sciences. Psychology; Humans; Intestinal Mucosa - microbiology; intestines; Microbiology; patients; Polymerase Chain Reaction - methods; qRT-PCR; quantitative polymerase chain reaction; reverse transcriptase polymerase chain reaction; R2; CD; B; E; qRT-PCR; Adherent-invasive E. coli; Crohn's disease; GFP; Ct; ECN; α; CFU; CV; Bacterial quantification; AIEC; IB; Human; Digestive system; Mucosa; Gut; Adhesion; Inflammatory disease; Polymerase chain reaction; Crohn disease; Digestive diseases; Intestinal disease; Bacteria; Quantitative analysis
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/j.mimet.2011.04.002

We here present a method based on qRT-PCR to quantify E. coli LF82 in intestinal human samples. Two different primer-probe sets were designed to detect LF82, and a third to target total E. coli. The assay showed high robustness and specificity for detection of LF82 in the presence of intestinal tissue.