Quantitative PCR method for sensitive detection of ruminant fecal pollution in freshwater and evaluation of this method in alpine karstic regions.
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 10(9) 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 10(5) marker equivalents per liter).
Reischer
Georg H
GH
Institute for Chemical Engineering, Gene Technology Group, Vienna University of Technology, Getreidemarkt 9-166/5, A-1060 Vienna, Austria.
Kasper
David C
DC
Steinborn
Ralf
R
Mach
Robert L
RL
Farnleitner
Andreas H
AH
D000818:Animals; D041963:Bacteroidetes / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005243:Feces / Q000382:microbiology*; D005618:Fresh Water / Q000382:microbiology*; D005819:Genetic Markers; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012418:Ruminants / Q000382:microbiology*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014876:Water Pollution; D014881:Water Supply
D005819:Genetic Markers