Article Text

MICRO DETECTION AND IDENTIFY OF HOST-BACTERIAL INTERACTION ON INTESTINAL TISSUE SAMPLES FROM NEONATES WITH NECROTIZING ENTEROCOLITIS
  1. B Smith1,
  2. L Molbak3,
  3. B L Petersen3,
  4. S Bod3,
  5. K A Krogfelt1,
  6. M Boye3
  1. 1Department of Mycology, Bacteriology and Paracitology, Statens Serum Institute, Copenhagen, Denmark
  2. 2Department of Neonatology, Rigshospitalet, Copenhagen, Denmark
  3. 3National Veterinary Institute-DTU, Technical University of Denmark, Copenhagen, Denmark

Abstract

In this study we investigated the microbial composition from neonates with necrotizing enterocolitis (NEC) Bell stadium IV. NEC is a serious bacterial infection in the intestine. It is not known if or which bacteria are responsible and which other factors triggers the infection in the intestine, but immaturity, premature birth and the diet are known to play a key role. In situ hybridization with a general bacterial 16S rRNA-targeting probe was used on 24 neonate’s tissue samples to visualize all bacteria in the borderline of the surgical removed formalin-fixed necrotic samples followed by Lacer Capture Microdissection (LCM). LCM offers the possibility to recover specific cells of interest from fixed intestinal tissue samples for downstream extraction of DNA and RNA from the host and the bacterial population. The isolated DNA was extracted and the relative number of bacteria was measured using a quantitative real-time PCR assay. A 16S rDNA clone library from 10 neonate was created, and the primarily data showed that the most frequent clones belong to Enterobacteria and Enterococcus. There were surprisingly few Lactobacillus and Clostridia clones and less than 0.1% of the clones belonged to the Bacteroidetes group. Instead many clones were found to belong to Actinobacteria, but none of these clones was Bifidobacterium.

Conclusion This study may show an uncommon genus of bacteria always represented in the necrotic tissue, and find the likelihood of many of the potential pathogenic bacteria found in culturing studies and in animal models as being responsible for NEC in infants.

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