Novel Approach for Isolation and Identification of Porcine Epidemic Diarrhea Virus (PEDV) Strain NJ Using Porcine Intestinal Epithelial Cells.
Porcine epidemic diarrhea virus (PEDV), which is the causative agent of porcine epidemic diarrhea in China and other countries, is responsible for serious economic losses in the pork industry. Inactivated PEDV vaccine plays a key role in controlling the prevalence of PEDV. However, consistently low viral titers are obtained during the propagation of PEDV in vitro; this represents a challenge to molecular analyses of the virus and vaccine development. In this study, we successfully isolated a PEDV isolate (strain NJ) from clinical samples collected during a recent outbreak of diarrhea in piglets in China, using porcine intestinal epithelial cells (IEC). We found that the isolate was better adapted to growth in IECs than in Vero cells, and the titer of the IEC cultures was 104.5 TCID50/0.1 mL at passage 45. Mutations in the S protein increased with the viral passage and the mutations tended towards attenuation. Viral challenge showed that the survival of IEC-adapted cultures was higher at the 45th passage than at the 5th passage. The use of IECs to isolate and propagate PEDV provides an effective approach for laboratory-based diagnosis of PEDV, as well as studies of the epidemiological characteristics and molecular biology of this virus.
['Shi|Wen|W|', 'Jia|Shuo|S|', 'Zhao|Haiyuan|H|', 'Yin|Jiyuan|J|', 'Wang|Xiaona|X|', 'Yu|Meiling|M|', 'Ma|Sunting|S|', 'Wu|Yang|Y|', 'Chen|Ying|Y|', 'Fan|Wenlu|W|', 'Xu|Yigang|Y|', 'Li|Yijing|Y|']
D000220:Adaptation, Biological D000818:Animals D002681:China D018352:Coronavirus Infections D004847:Epithelial Cells D053485:Porcine epidemic diarrhea virus D012692:Serial Passage D013552:Swine D013553:Swine Diseases D019562:Viral Load D014776:Virus Cultivation
isolation and identification porcine intestinal epithelial cells porcine epidemic diarrhea virus (PEDV) NJ strain