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Use of denaturing gradient gel electrophoresis to detect Actinobacteria associated with the human faecal microbiota

Hoyles, L., Clear, J. A. and McCartney, A. L. (2013) Use of denaturing gradient gel electrophoresis to detect Actinobacteria associated with the human faecal microbiota. Anaerobe, 22. pp. 90-96. ISSN 1075-9964

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To link to this item DOI: 10.1016/j.anaerobe.2013.06.001

Abstract/Summary

With the exceptions of the bifidobacteria, propionibacteria and coriobacteria, the Actinobacteria associated with the human gastrointestinal tract have received little attention. This has been due to the seeming absence of these bacteria from most clone libraries. In addition, many of these bacteria have fastidious growth and atmospheric requirements. A recent cultivation-based study has shown that the Actinobacteria of the human gut may be more diverse than previously thought. The aim of this study was to develop a denaturing gradient gel electrophoresis (DGGE) approach for characterizing Actinobacteria present in faecal samples. Amount of DNA added to the Actinobacteria-specific PCR used to generate strong PCR products of equal intenstity from faecal samples of five infants, nine adults and eight elderly adults was anti-correlated with counts of bacteria obtained using fluorescence in situ hybridization probe HGC69A. A nested PCR using Actinobacteria-specific and universal PCR-DGGE primers was used to generate profiles for the Actinobacteria. Cloning of sequences from the DGGE bands confirmed the specificity of the Actinobacteria-specific primers. In addition to members of the genus Bifidobacterium, species belonging to the genera Propionibacterium, Microbacterium, Brevibacterium, Actinomyces and Corynebacterium were found to be part of the faecal microbiota of healthy humans.

Item Type:Article
Refereed:Yes
Divisions:Faculty of Life Sciences > School of Chemistry, Food and Pharmacy > Department of Food and Nutritional Sciences > Food Microbial Sciences Research Group
ID Code:32912
Uncontrolled Keywords:Bifidobacterium; Corynebacterium; Actinomyces; Gut microbiota; DGGE; Ecology
Publisher:Elsevier

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