The objective of this work was to develop an assay for detection of enteroviruses and evaluate the applicability of the new system for environmental water samples. Primers and probe used for this assay were designed using the Primer Express software from PE Applied Biosystems. Standard curves were generated using different RNA stocks. Following purification, the nucleic acid was quantitated by UV spectroscopy. A dilution series of each RNA stock was amplified using the 7700 Sequence Detector and standard curves calculated. The R squared value for each curve was greater than 0.99 and found to be consistent and reproducible. This method has been successfully used to quantitate virus amplification in cell culture and to measure the PCR inhibition caused by debris in groundwater concentrates. Includes tables, figure.