Cell culture isolation and sequence analysis of genetically diverse US porcine epidemic diarrhea virus strains including a novel strain with a large deletion in the spike gene

- We isolated a novel PEDV strain with a large deletion (197 aa) in the spike gene.
- We isolated US PEDV INDEL variant in Vero cells.
- We isolated multiple original highly virulent US PEDV strains in Vero cells.
- Genomic sequences of the isolated PEDV strains were analyzed.
- The isolated genetically diverse PEDV strains can be used for attenuated vaccine development.

The highly contagious and deadly porcine epidemic diarrhea virus (PEDV) first appeared in the US in April 2013. Since then the virus has spread rapidly nationwide and to Canada and Mexico causing high mortality among nursing piglets and significant economic losses. Currently there are no efficacious preventive measures or therapeutic tools to control PEDV in the US. The isolation of PEDV in cell culture is the first step toward the development of an attenuated vaccine, to study the biology of PEDV and to develop in vitro PEDV immunoassays, inactivation assays and screen for PEDV antivirals. In this study, nine of 88 US PEDV strains were isolated successfully on Vero cells with supplemental trypsin and subjected to genomic sequence analysis. They differed genetically mainly in the N-terminal S protein region as follows: (1) strains (n = 7) similar to the highly virulent US PEDV strains; (2) one similar to the reportedly US S INDEL PEDV strain; and (3) one novel strain most closely related to highly virulent US PEDV strains, but with a large (197 aa) deletion in the S protein. Representative strains of these three genetic groups were passaged serially and grew to titers of ∼5-6 log10 plaque forming units/mL. To our knowledge, this is the first report of the isolation in cell culture of an S INDEL PEDV strain and a PEDV strain with a large (197 aa) deletion in the S protein. We also designed primer sets to detect these genetically diverse US PEDV strains.