Direct Quantification of the Enteric Bacterium Oxalobacter formigenes in Human Fecal Samples by Quantitative Competitive-Template PCR

Author:

Sidhu H.1,Holmes R. P.2,Allison M. J.3,Peck A. B.4

Affiliation:

1. Ixion Biotechnology, Inc., Alachua,1 and

2. Department of Urology, Bowman Gray School of Medicine, Winston-Salem, North Carolina2; and

3. National Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Ames, Iowa3

4. Department of Pathology, Immunology & Laboratory Medicine, University of Florida, Gainesville,4 Florida;

Abstract

ABSTRACT Homeostasis of oxalic acid appears to be regulated, in part, by the gut-associated bacterium Oxalobacter formigenes . The loss of this bacterium from the gut flora is associated with an increased susceptibility to hyperoxaluria, a condition which can lead to the formation of calcium oxalate crystalluria and kidney stones. In order to identify and quantify the presence of O. formigenes in clinical specimens, a quantitative-PCR-based assay system utilizing a competitive DNA template as an internal standard was developed. This quantitative competitive-template PCR test allows for the rapid, highly specific, and reproducible quantification of O. formigenes in fecal samples and provides a prototype for development of DNA-based quantitative assays for enteric bacteria.

Publisher

American Society for Microbiology

Subject

Microbiology (medical)

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