DETECTION OF HEPATITIS-A VIRUS, ROTAVIRUS, AND ENTEROVIRUS IN NATURALLY CONTAMINATED SHELLFISH AND SEDIMENT BY REVERSE TRANSCRIPTION-SEMINESTED PCR

A reverse transcription-PCR method was developed to detect enterovirus (EV), hepatitis A virus (HAV), and rotavirus (RV) RNAs in shellfish a nd sediment. The method was first tested under experimental conditions by using virus-spiked shellfish to evaluate assay sensitivity. The us e of CC41 cellulose was found to be efficient for removing inhibitors of RV detection. For sediment samples, a Sephadex column was used to a llow the detection of EV and HAV RNAs. The specificity of amplified pr oducts was controlled by hybridization with digoxigenin-labeled oligop robes. The method was then applied to naturally contaminated shellfish and sediments. EV, HAV, and RV RNAs were detected in 22, 14, and 20% of the shellfish Samples, respectively. No relationship between viral contamination and bacterial contamination was found. When viral RNAs ( HAV or EV) were detected in sediments, they were also detected in shel lfish.