An Efficient, Organic Solvent-Free Method for Extraction and Concentration of Hepatitis E Virus from Pig Liver

Abstract

The presence of the hepatitis E virus (HEV) in pork products, particularly in pig liver has been frequently described. However, a standardized method is not still available for the detection of HEV in foods, particularly in those difficult food matrixes such as pig meat and pork products. The aim of this study was to design, optimize and evaluate a new method of food-separation and virus concentration for HEV in pig liver samples. This method is based on organic flocculation and avoids the use of organic solvents. The virus recovery rates and analytical sensitivity of the method using murine norovirus MNV-1 as a surrogate were 73.6–82.2% and at least 1 × 103 TCID50 per g of liver in 100% of the replicates, respectively. Furthermore, this new methodology was validated by testing the presence of HEV RNA in naturally infected pig liver samples, comparing it with two other commonly used concentration methods. The new extraction method run satisfactory in comparison with the two reference methods; statistically equivalent (p < 0.05) to one of the methods used while presented statistically significant better results (p < 0.05) compared to the second method. Consequently, our results indicate that the new extraction method can be an adequate cost-effective and ecologically friendly alternative for the food separation and concentration of HEV RNA in pig liver samples.