|Ley, Victoria - NAT'L INSTITUTE, MADRID|
Submitted to: International Congress of Virology
Publication Type: Abstract Only
Publication Acceptance Date: June 10, 2002
Publication Date: July 26, 2002
Citation: Perdue, M.L., Ley, V., Higgins, J.A., Fayer, R. 2002. A real-time RT-PCR reaction specific for bovine enteroviruses provides a rapid assay for fecal contamination of the environment by cattle [Abstract]. International Congress of Virology, July 23-26, 2002, Paris, France. Technical Abstract: Characterization of bovine enteroviruses found in a closed herd of cattle and the surrounding environment indicated that bovine enteroviruses (BEV) could serve as useful markers of contamination with cattle waste. BEV was found in feces from 76% of cattle, 38% of white-tail deer in the same area and in Canada geese on the farm. Virus was cultured from water obtained from drinking water tanks, standing water in the pasture, stream water draining the pasture to the river and river water as well. Furthermore, BEV was found in oysters from a bar in the river downstream from the farm. In order to rapidly identify the virus in the environment, a set of real-time PCR primers and probes were developed that would detect both BEV-1 and BEV-2 strains of virus and that could be used in a portable format on the latest generation of portable real-time PCR machines. The primer set amplified part of the 5' non-coding region of the genome and the probes were labeled with FAM and TAMRA for detection of positive reactions. The primer/probes readily detected cultured BEV and at least some shellfish gill wash samples directly without culture. The sensitivity of the probes is being assayed using commercially obtained shellfish samples and comparing with culture positives, but initial data indicate the primer/probes may be used as portable rapid reagents to detect waste contamination levels in a variety of environmental samples.