Bacterial colonization of colonic crypt mucous gel and disease activity in ulcerative colitis.
Rowan, Fiachra Docherty, Neil G Murphy, Madeline Murphy, T Brendan Coffey, J Calvin O'Connell, P Ronan
Rowan, Fiachra
Docherty, Neil G
Murphy, Madeline
Murphy, T Brendan
Coffey, J Calvin
O'Connell, P Ronan
Advisors
Editors
Other Contributors
Date
2012-02-01T10:31:25Z
Date Submitted
Keywords
Other Subjects
Subject Mesh
Adult
Aged
Analysis of Variance
Area Under Curve
Case-Control Studies
Colitis, Ulcerative/*microbiology/pathology
DNA, Bacterial/analysis
Desulfovibrio/*isolation & purification
Female
Humans
Intestinal Mucosa/*microbiology/pathology
Lasers
Male
Microdissection/methods
Middle Aged
Mucus/microbiology
Polymerase Chain Reaction
RNA, Ribosomal/analysis
Sensitivity and Specificity
Statistics, Nonparametric
Aged
Analysis of Variance
Area Under Curve
Case-Control Studies
Colitis, Ulcerative/*microbiology/pathology
DNA, Bacterial/analysis
Desulfovibrio/*isolation & purification
Female
Humans
Intestinal Mucosa/*microbiology/pathology
Lasers
Male
Microdissection/methods
Middle Aged
Mucus/microbiology
Polymerase Chain Reaction
RNA, Ribosomal/analysis
Sensitivity and Specificity
Statistics, Nonparametric
Planned Date
Start Date
Collaborators
Principal Investigators
Alternative Titles
Publisher
Abstract
OBJECTIVE: To optimize total bacterial 16S rRNA quantification in microdissected colonic crypts in healthy controls and patients with ulcerative colitis (UC) and to characterize the findings with disease activity. BACKGROUND: Microscopic and molecular techniques have recently converged to allow bacterial enumeration in remote anatomic locations [eg, crypt-associated mucous gel (CAMG)]. The aims of this study were to combine laser capture microdissection (LCM) and 16S rRNA-based quantitative polymerase chain reaction (qPCR) to determine total bacterial copy number in CAMG both in health and in UC and to characterize the findings with disease activity. METHODS: LCM was used to microdissect CAMG from colonic mucosal biopsies from controls (n = 20) and patients with acute (n = 10) or subacute (n = 10) UC. Pan-bacterial 16S rRNA copy number per millimeter square in samples from 6 locations across the large bowel was obtained by qPCR using Desulfovibrio desulfuricans as a reference strain. Copy numbers were correlated with the UC disease activity index (UCDAI) and the simple clinical colitis activity index (SCCAI). RESULTS: Bacterial colonization of CAMG was detectable in all groups. Copy numbers were significantly reduced in acute UC. In subacute colitis, there was a positive correlation between copy number and UCDAI and SCCAI in the ascending, transverse and sigmoid colon. CONCLUSIONS: This study describes a sensitive method of quantitatively assessing bacterial colonization of the colonic CAMG. A positive correlation was found between CAMG bacterial load and subacute disease activity in UC, whereas detectable bacterial load was reduced in acute UC.
Language
eng
Citation
ISSN
1528-1140 (Electronic)
0003-4932 (Linking)
0003-4932 (Linking)
eISSN
ISBN
DOI
10.1097/SLA.0b013e3181fdc54c
PMID
21037444