Simple method of detecting enteroviruses in contaminated molluscs and sewage by using polymerase chain reaction coupled with a colorimetric microwell detection assay

The methods normally used for the detection of enteroviruses in environmental samples involve the use of cell cultures, which are expensive and time consuming. The Polymerase Chain Reaction PCR. is a useful tool for the detection of enteroviruses in several matrixes because primary cell culture is not needed and the increased sensitivity of PCR allows detection of the low numbers of target nucleic acids usually found in environmental samples. A 5-h, user-friendly PCR assay was used to detect enteroviruses in bivalves molluscs clams. and sewage. Reverse transcription and amplification were performed in a one-step reaction using rTth polymerase. Carryover contamination was prevented with dUTP and uracil N-glycosylase. Detection was performed colorimetrically in a microwell titer plate. This method has greater advantages over conventional methodologies for routinely screening a large number of samples, namely, the rapid acquisition of results and cost effectiveness.