Recovery of Poliovirus Type 1 from Experimentally Contaminated Shellfish: Evaluation of Different Methods

Abstract

Outbreaks of viral gastroenteritis caused by ingestion of contaminated shellfish are well documented. Several methods have been described for virus recovery from shellfish, but often they are complicated and time-consuming. In this study we have compared several viral extraction-concentration methods from mussels experimentally contaminated in conditions close to those met in natural environment. Mvtilus galloprovincialis were contaminated by 2 h immersion in artificial seawater seeded with Poliovirus type 1. After the contamination step, the mussel bodies with intervalve water were homogenized by Ultraturax blender using four different elution buffers: PBS pH9.5; 0.05M glycine pH9.5; 0.3% and 3% beef extract pH9.5. The ratio between eluants and homogenates, depending on the used method, was 1:2 or 1:10. After the elution step, virus was concentrated comparing the following techniques: precipitation at acid pH, precipitation with PEG6000, ultracentrifugation, ultrafiltration with Amicon apparatus and tangential flow Minitan system. Statistical analysis was performed using ANOVA program and the results were evaluated.