Isolation and Propagation of Porcine Deltacoronavirus in Cell Culture and Embryonated Chicken Eggs

Porcine deltacoronavirus (PDCoV) is a novel porcine enteropathogenic coronavirus that can cause acute watery diarrhea/vomiting, dehydration, and mortality in piglets. Since the first outbreak of PDCoV in swine herds with diarrhea in the United States in 2014, its prevalence was subsequently reported in many countries, leading to huge economic losses for pig farms. PDCoV has the capacity to infect cells from broad species including porcine, human, calf, and chicken cells. It also has limited infectivity in calves, chickens, and mice, posing considerable threat to animals and human health. PDCoV isolation and propagation have been done using embryonated chicken eggs and swine cell lines (swine testicular [ST] and LLC porcine kidney [LLC-PK] cells). Moreover, cell culture-adapted PDCoVs were propagated on porcine small intestinal epithelial (IPEC-J2) cells and bovine primary mesenchymal cells. This chapter mainly provides detailed protocols for the isolation and propagation of PDCoV in LLC-PK and ST cells, respectively. The procedures of PDCoV propagation using embryonated chicken eggs were also described. The methods used for confirming the virus isolation results were included.