Quantitation, biological and physicochemical properties of cell culture-adapted porcine epidemic diarrhea coronavirus (PEDV)

Abstract

The porcine epidemic coronavirus (PEDV), tentatively classified as a coronavirus, was adapted to Vero cells and a plaque test developed for infectivity titration, allowing us to test the biological and biophysical properties of the virus. Growth kinetics showed peak titers of 10(5.5) plaque-forming units ml-1 15 h after infection. Filtration experiments and electron microscopy revealed a particle diameter between 100 and 200 nm. The buoyant density of the virus was 1.18. The particle lost its infectivity on treatment with lipid solvents. Virus replication could not be inhibited by 5-iodo-2'-deoxyuridine. PEDV was moderately stable at 50 degrees C, but heat sensitivity was not altered by divalent cations. At 4 degrees C, the virus was stable between pH 5.0 and 9.0, but at 37 degrees C stability was restricted to the pH range 6.5-7.5. Viral infectivity was not impaired by ultrasonication or by multiple freezing and thawing. PEDV was not neutralized by transmissible gastroenteritis virus antiserum. On the basis of the tests carried out, PEDV is a pleomorphic, enveloped RNA virus with a particle diameter of approximately 150 nm and a buoyant density of 1.18. Infectivity depends on the presence of trypsin, and infected cells show a tendency to fuse and to form syncytia. All of these properties, as well as its physicochemical characteristics, allow PEDV to be classified as a coronavirus.

Study on inactivation of porcine epidemic diarrhoea virus, porcine sapelovirus 1 and adenovirus in the production and storage of laboratory spray‐dried porcine plasma

“…). As reported for other Coronaviridae this suggests that the surface structure of these PEDV particles stays intact in environments other than faeces (Hofmann and Wyler ; Lai et al . ).…”

Identification of two novel B cell epitopes on porcine epidemic diarrhea virus spike protein

“…Reactivity of the McAbs with the native S protein was tested by ELISA and Western blotting. Neutralizing activity of the McAbs was tested according to the methods described by Hofmann and Wyler (1989).…”

“…In ELISA, antisera of the epitopes S1D5 and S1D6 were diluted 100 times with PBS for detection, other procedures were as for ELISA of the recombinant proteins (Section 2.6). Neutralizing activity of antibodies against the epitopes S1D5 and S1D6 was tested according to the methods described by Hofmann and Wyler (1989). Note: (+) sign denotes sense strands and (À) sign denotes anti-sense strands.…”

Impact of porcine epidemic diarrhea on herd and individual Berkshire sow productivity

“…Porcine epidemic diarrhea (PED) is caused by PED virus, an enveloped single-stranded RNA virus in the family Coronaviridae that is related to transmissible gas-troenteritis coronavirus (Pensaert and DeBouck, 1978;Hofmann and Wyler, 1989). The PED virus emerged as a global threat to the swine industry in 2013, when a number of epidemics were reported in many important swineproducing countries in North America and East Asia that were previously believed to be PED virus-free (Mole, 2013;Stevenson et al, 2013;Chen et al, 2014;Park et al, 2014;Hanke et al, 2015;Song et al, 2015).…”