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Applied and Environmental Microbiology, August 2006, p. 5610-5614, Vol. 72, No. 8
0099-2240/06/$08.00+0     doi:10.1128/AEM.00364-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

Quantitative PCR Method for Sensitive Detection of Ruminant Fecal Pollution in Freshwater and Evaluation of This Method in Alpine Karstic Regions

Georg H. Reischer,1 David C. Kasper,1 Ralf Steinborn,2 Robert L. Mach,1 and Andreas H. Farnleitner1*

Institute for Chemical Engineering, Gene Technology Group, Vienna University of Technology, Getreidemarkt 9/166, A-1060 Vienna, Austria,1 Institute of Animal Breeding and Genetics, Department for Animal Breeding and Reproduction, University of Veterinary Medicine, Veterinaerplatz 1, A-1210 Vienna, Austria2

Received 14 February 2006/ Accepted 8 May 2006

A quantitative TaqMan minor-groove binder real-time PCR assay was developed for the sensitive detection of a ruminant-specific genetic marker in fecal members of the phylum Bacteroidetes. The qualitative and quantitative detection limits determined were 6 and 20 marker copies per PCR, respectively. Tested ruminant feces contained an average of 4.1 x 109 marker equivalents per g, allowing the detection of 1.7 ng of feces per filter in fecal suspensions. The marker was detected in water samples from a karstic catchment area at levels matching a gradient from negligible to considerable ruminant fecal influence (from not detectable to 105 marker equivalents per liter).

* Corresponding author. Mailing address: Institute for Chemical Engineering, Gene Technology Group, Vienna University of Technology, Getreidemarkt 9-166/5, A-1060 Vienna, Austria. Phone: 43-1-58801-17251. Fax: 43-1-5816266. E-mail: A.Farnleitner{at}aon.at.

Applied and Environmental Microbiology, August 2006, p. 5610-5614, Vol. 72, No. 8
0099-2240/06/$08.00+0     doi:10.1128/AEM.00364-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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