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J Appl Microbiol. 2009 Feb;106(2):634-41. doi: 10.1111/j.1365-2672.2008.04037.x.

Development of Faecalibacterium 16S rRNA gene marker for identification of human faeces.

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1
Department of Veterinary Pathobiology, University of Missouri, Columbia, MO, USA.

Abstract

AIMS:

The focus of this study was to identify a bacterial 16S rRNA gene sequence, unique to microbiota in the human gut, for use in development of a dependable PCR assay to detect human faecal pollution in water.

METHODS AND RESULTS:

Suppression subtractive hybridization (SSH) and bioinformatics were used to identify a genetic marker, within the 16S rRNA gene of Faecalibacterium, for the detection of human faeces. DNA sequencing analysis demonstrated that a majority (16) of 74 clones of the SSH library contained insertion sequences identified as Faecalibacterium 16S rRNA genes. Human faeces-specific sequences were derived and six PCR primer sets designed and tested against faecal DNA samples from human and nonhuman sources. One PCR primer set, HFB-F3 and HFB-R5, was exclusively associated with human faeces. These primers generated a human faeces-specific amplicon of 399 bp from 60.2% of human faecal samples and 100% of sewage samples.

CONCLUSIONS:

The subject Faecalibacterium marker is specific for sewage.

SIGNIFICANCE AND IMPACT OF THE STUDY:

This study represents the initial report of a Faecalibacterium marker for human faeces, which may prove useful for microbial source tracking.

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