Porcine epidemic diarrhea virus purification using low-speed centrifugation

Abstract

Porcine epidemic diarrhea virus (PEDV) is a porcine coronavirus that causes enteric diseases characterized by watery diarrhea and dehydration in suckling piglets. Concentrated and highly purified viruses are required for the preparation of vaccines, diagnostics, and virus research. Currently, most protocols for virus purification require ultracentrifugation, which can be an instrumental barrier to routine operations in a laboratory. In this study, the efficacy of low-speed centrifugation for virus concentration was examined. The SM98 strain of PEDV was propagated in Vero cells and pelleted by centrifugation for 3 h at high speed (100,000 × g) or for 18 h at low speed (10,000 × g). The efficacy of virus concentration was analyzed by virus titration and western blotting. The amounts of infectious viruses and viral proteins in the pelleted samples obtained by low-speed centrifugation were comparable to those obtained by high-speed centrifugation. Interestingly, the pelleted sample impurity level was lower in low-speed than in high-speed centrifugation. In summary, we describe an efficient, easy-to-perform protocol for the preparation of purified and concentrated PEDV.