Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR

摘要: Shellfish are readily contaminated with viruses present in water containing sewage due to the concentrating effect of filter feeding. Enteroviruses generally used as a model for detection from shellfish their public health significance. In work, oysters were placed glass aquaria seawater plus unicellular algae. Two experiments performed: 1) bioaccumulating four different poliovirus type 2 concentrations: 5 x 104, 2.5 103 and 102 PFU/mL during 20h; 2) oyster tissues directly inoculated 6.0 105 1.0 PFU/mL. After seeding, tissue samples processed by an adsorption-elution-precipitation method. Positive controls performed seeding on final extracts. Oyster extracts assayed recovery plaque assay, RT-PCR integrated cell culture-PCR methodologies (ICC/PCR). The last one was based inoculation onto VERO monolayer followed analysis infected fluid. first experiment (20h bioaccumulation) until PFU detected after 24 48h growth cells. Direct ICC/PCR able detect 3 0.04 poliovirus, respectively, when bioaccumulation assay used. When direct virus performed, assays showed that polioviruses recovered all tested concentrations. Based these results, it is possible conclude viable can be techniques applied monitoring contamination environmental samples.