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.

Porcine epidemic diarrhea virus (PEDV) infects all-age pigs and causes enteric diseases. Genetic diversities in isolates been reported from each country, and those diversities highlighted in pathogenicity and vaccine. In this manuscript, we are reporting of new PEDV isolation in Korea, and with genetic characteristics. Our new isolate belongs to G2b and put the name as CNUP6-2018.

Viral particles of Porcine epidemic diarrhea virus (PEDV) consist of a four structural proteins. Among them Spikeprotein mediated responsible for receptor binding and membrane fusion during viral infection and therefore the main targetof neutralizing antibodies. Virus-like particles (VLPs) are consisted of one or more viral structural proteins, and theirmorphologies closely resemble those of the native virus. VLPs have no virulence and can elicit robust immune responsesas compared with inactivated or live-attenuated virus vaccines. Thus, in this study, we tried two methods for VLP constructionin Bombyx mori, one is traditional method and the other is chimeric VLP method using the influenza matrix protein.Both methods could produce successfully PEDV VLPs.

The coronavirus porcine epidemic diarrhea virus (PEDV) infects the cells lining the small intestine of a pig and, causes porcine epidemic diarrhea (PED). Owing to its highly infectious nature, PEDV has a substantial economic burden, which results in significant morbidity and mortality in piglets. In this study, the virucidal efficacy of a powder disinfectant containing a phosphate compound against PEDV was investigated. Virucidal efficacy was assessed as the infectivity of PEDV toward Vero cells after exposure of the virus to the disinfectant. PEDV was exposed to the disinfectant in the presence of either hard water (HW) or an organic matter suspension (OM). In the HW condition, PEDV was inactivated by 4-fold dilution of the disinfectant. In the presence of OM, the disinfectant showed virucidal activity with a 2-fold dilution. As the disinfectant possessed virucidal activity against PEDV, it should be an effective reagent for limiting the spread of animal viral diseases.

Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea and dehydration in pigs with high mortality. PEDV is belong to Coronavirus, enveloped, single-stranded, positive-sense RNA virus. PEDV particles were composed of four structure proteins such as a glycosylated peplomer (spike, S) protein, envelope (E), glycosylated membrane (M) protein, and unglycosylated RNA-binding nucleocapsid (N) protein. Many of previous studies talk about this four structure proteins have a great potential to diagnosis and prevent PEDV. In this study we investigated expression of these structure proteins using the bacterial and baculovirus expression system. In bacterial expression system, our results showed that structure proteins fused polyhedrin and intein gene were expressed higher than non-fusion structure proteins. The expressed fusion proteins were used to immune mice for generating a polyclonal antibodies. In baculovirus expression system, co-infection of insect cells with these four recombinant baculoviruses led to self-assembly of virus-like particles as demonstrated by Transmission electron microscopy. They were confirmed by western blot analysis using pre-made polyclonal antibodies. Finding in this study may provide important information for vaccine and diagnostic development.

Porcine epidemic diarrhea virus (PEDV) causes porcine epidemic diarrhea (PED) and a considerable economic loss in the swine industry. In this study, the virucidal efficacy of a disinfectant composed of citric acid, benzalkonium chloride and phosphoric acid against PEDV was investigated. Virucidal efficacy was assessed as the infectivity of PEDV toward Vero cells after exposure of the virus to the disinfectant. PEDV was exposed to the disinfectant in the presence of either hard water (HW) or an organic matter suspension (OM). In HW condition, PEDV was inactivated by 600-fold dilutions of the disinfectant. In the presence of OM, the disinfectant showed virucidal activity after a 200-fold dilution. As the disinfectant possesses virucidal activity against PEDV, it should be an effective reagent to use to limit the spread of animal viral diseases.

During 2008 2010, 943 swine sera were collected from 45 farms located nationwide. Antibodies against porcine epidemic diarrhea virus (PEDV) were tested via serum neutralization antibody test (SNT) using PEDV-SN, which was adapted and propagated on the Vero cell monolayer with trypsin-free culture media supplemented with more than 10% fetal bovine serum (FBS). All 45 farms were shown to have at least one or more seropositive pig. Of the 943 swine sera that were tested, 931 sera were neutralizing antibody positive against PEDV. These high seroprevalence rates seemed to be due to vaccination or natural infection of PEDV. In a plaque reduction neutralization test (PRNT) using a swine serum showing SN titer of 1:32, a greater than 50% plaque reduction was observed in up to 160 times serum dilution.

This study was carried out to obtain basic information for possibility of oral vaccine in carrot using Agrobacteruim -mediated transformation system. The epitope region of porcine epidemic diarrhea virus (PEDV) spike gene which is classified as a member of the Coronaviridae and causes an acute enteritis in pigs was successfully expressed in carrot (Daucus carota) using the Agrobacterium-mediated transformation system. Hypocotyl segments of in vitro germinated plantlets were infected with Agrobacteriun tumefaciens LBA 4404 harboring PEDV spike gene. Embryogenic callus (EC) was induced on MS selection medium with 1 mg/L 2,4-D, 50 mg/L kanamycin and 300 mg/L cefotaxime after 45 days of culture. Subcultured ECs on MS selection medium without 2,4-D were converted to somatic embryos (SE) of various stage; globular, heart and torpedo stage. Putative transgenic embryos were selected on MS medium with 50 mg/L kanamycin and 300 mg/L cefotaxime. Regenerated plantlets from transformed SE were induced on MS medium containing 50 mg/L kanamycin after 30 days of culture. Genomic PCR confirmed the integration of PEDV spike gene into nuclear genome of carrot and northern blot analysis demonstrated the expression of PEDV spike gene in transgenic carrot.