identification and characterisation of pseudomonas 16s ribosomal dna from ileal biopsies of children with crohns disease识别和描述的假单胞菌16 s核糖体dna回肠活检与克罗恩病的孩子.pdfVIP

Identification and Characterisation of Pseudomonas 16S Ribosomal DNA from Ileal Biopsies of Children with Crohn’s Disease 1 2 3 3 1 Josef Wagner , Kirsty Short , Anthony G. Catto-Smith , Don J. S. Cameron , Ruth F. Bishop , Carl D. Kirkwood1* 1 Enteric Virus Group, Murdoch Childrens Research Institute, Department of Paediatrics, Royal Children’s Hospital, Parkville, Victoria, Australia, 2 Department of Microbiology Immunology, University of Melbourne, Melbourne, Victoria, Australia, 3 Department of Gastroenterology Clinical Nutrition, Royal Children’s Hospital, Parkville, Victoria, Australia Abstract Molecular analysis of bacterial 16S rRNA genes has made a significant contribution to the identification and characterisation of bacterial flora in the human gut. In particular, this methodology has helped characterise bacterial families implicated in the aetiology of inflammatory bowel disease (IBD). In this study we have used a genus specific bacterial 16S PCR to investigate the prevalence and diversity of Pseudomonas species derived from the ileum of children with Crohn’s disease (CD), and from control children with non-inflammatory bowel disease (non-IBD) undergoing their initial endoscopic examination. Fifty eight percent of CD patients (18/32) were positive using the Pseudomonas PCR, while significantly fewer children in the non-IBD group, 33% (12/36), were PCR positive for Pseudomonas (p,0.05, Fischer’s exact test). Pseudomonas specific 16S PCR products from 13 CD and 12 non-IBD children were cloned and sequenced. Five hundred and eighty one sequences were generated and used for the comparative analysis of Pseudomonas diversity between CD and non-IBD patients. Pseudomonas species