Bacterial colonization of colonic crypt mucous gel and disease activity in ulcerative colitis.

Hdl Handle:
http://hdl.handle.net/10147/207562
Title:
Bacterial colonization of colonic crypt mucous gel and disease activity in ulcerative colitis.
Authors:
Rowan, Fiachra; Docherty, Neil G; Murphy, Madeline; Murphy, T Brendan; Coffey, J Calvin; O'Connell, P Ronan
Affiliation:
Department of Surgery, St. Vincent's University Hospital, UCD School of Medicine , and Medical Science, Dublin, Ireland.
Citation:
Ann Surg. 2010 Nov;252(5):869-75.
Journal:
Annals of surgery
Issue Date:
1-Feb-2012
URI:
http://hdl.handle.net/10147/207562
DOI:
10.1097/SLA.0b013e3181fdc54c
PubMed ID:
21037444
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
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
ISSN:
1528-1140 (Electronic); 0003-4932 (Linking)

Full metadata record

DC FieldValue Language
dc.contributor.authorRowan, Fiachraen_GB
dc.contributor.authorDocherty, Neil Gen_GB
dc.contributor.authorMurphy, Madelineen_GB
dc.contributor.authorMurphy, T Brendanen_GB
dc.contributor.authorCoffey, J Calvinen_GB
dc.contributor.authorO'Connell, P Ronanen_GB
dc.date.accessioned2012-02-01T10:31:25Z-
dc.date.available2012-02-01T10:31:25Z-
dc.date.issued2012-02-01T10:31:25Z-
dc.identifier.citationAnn Surg. 2010 Nov;252(5):869-75.en_GB
dc.identifier.issn1528-1140 (Electronic)en_GB
dc.identifier.issn0003-4932 (Linking)en_GB
dc.identifier.pmid21037444en_GB
dc.identifier.doi10.1097/SLA.0b013e3181fdc54cen_GB
dc.identifier.urihttp://hdl.handle.net/10147/207562-
dc.description.abstractOBJECTIVE: 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.en_GB
dc.language.isoengen_GB
dc.subject.meshAdulten_GB
dc.subject.meshAgeden_GB
dc.subject.meshAnalysis of Varianceen_GB
dc.subject.meshArea Under Curveen_GB
dc.subject.meshCase-Control Studiesen_GB
dc.subject.meshColitis, Ulcerative/*microbiology/pathologyen_GB
dc.subject.meshDNA, Bacterial/analysisen_GB
dc.subject.meshDesulfovibrio/*isolation & purificationen_GB
dc.subject.meshFemaleen_GB
dc.subject.meshHumansen_GB
dc.subject.meshIntestinal Mucosa/*microbiology/pathologyen_GB
dc.subject.meshLasersen_GB
dc.subject.meshMaleen_GB
dc.subject.meshMicrodissection/methodsen_GB
dc.subject.meshMiddle Ageden_GB
dc.subject.meshMucus/microbiologyen_GB
dc.subject.meshPolymerase Chain Reactionen_GB
dc.subject.meshRNA, Ribosomal/analysisen_GB
dc.subject.meshSensitivity and Specificityen_GB
dc.subject.meshStatistics, Nonparametricen_GB
dc.titleBacterial colonization of colonic crypt mucous gel and disease activity in ulcerative colitis.en_GB
dc.contributor.departmentDepartment of Surgery, St. Vincent's University Hospital, UCD School of Medicine , and Medical Science, Dublin, Ireland.en_GB
dc.identifier.journalAnnals of surgeryen_GB
dc.description.provinceLeinster-

Related articles on PubMed

All Items in Lenus, The Irish Health Repository are protected by copyright, with all rights reserved, unless otherwise indicated.