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 (197aa) deletion in the S protein. Representative strains of these three genetic groups were passaged serially and grew to titers of ~5-6log10 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 (197aa) deletion in the S protein. We also designed primer sets to detect these genetically diverse US PEDV strains.
Bibliographical noteFunding Information:
Nucleotide sequencing and electron microscopy were performed at the Molecular and Cellular Imaging Center, the Ohio Agricultural Research and Development Center (OARDC), The Ohio State University. We thank Dr. Steven Lawson, South Dakota State University, for providing PEDV N protein MAb (#6-29). We thank Susan E. Sommer-Wagner, Xiaohong Wang, Hui Hu, Xiang Gao, Marcia V. Lee, Kelly A. Scheuer, Anamika Veeramani and Garrett T. Stoltzfus for technical assistance, Andrea Kaszas for assistance with electron microscopy, and Jody Whittier for sequencing. Salaries and research support were provided by state and federal funds appropriated to OARDC, The Ohio State University and by the University of Minnesota Veterinary Diagnostic Laboratory. This work was supported in part by a grant from the National Pork Board (no. 13-222 to Q.W. and L.J.S.).
- Cell culture isolation
- Complete genome analysis
- Large deletion in S protein
- PEDV variants
- Porcine epidemic diarrhea virus (PEDV)