title: Novel Approach for Isolation and Identification of Porcine Epidemic Diarrhea Virus (PEDV) Strain NJ Using Porcine Intestinal Epithelial Cells.
abstract: 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 104.5TCID50/0.1 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.
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. wenshi_china@163.com.
authors.forename: Wen
authors.initials: W
authors.lastname: Shi
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. jiashuo0508@163.com.
authors.forename: Shuo
authors.initials: S
authors.lastname: Jia
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. zhywxn1925@163.com.
authors.forename: Haiyuan
authors.initials: H
authors.lastname: Zhao
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. neauyjy@126.com.
authors.forename: Jiyuan
authors.initials: J
authors.lastname: Yin
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. xiaonawang0319@163.com.
authors.forename: Xiaona
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. yu19890130@126.com.
authors.forename: Meiling
authors.initials: M
authors.lastname: Yu
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. masunting@163.com.
authors.forename: Sunting
authors.initials: S
authors.lastname: Ma
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. wuyang_neau@126.com.
authors.forename: Yang
authors.initials: Y
authors.lastname: Wu
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. chenying_neau@163.com.
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. fanwenlu1230@163.com.
authors.forename: Wenlu
authors.initials: W
authors.lastname: Fan
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. yigangxu_china@sohu.com.
authors.forename: Yigang
authors.initials: Y
authors.lastname: Xu
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. yijingli@163.com.
authors.forename: Yijing
authors.initials: Y
authors.lastname: Li
mesh_terms: D000220:Adaptation, Biological; D000818:Animals; D002681:China; D018352:Coronavirus Infections / Q000662:veterinary* / Q000821:virology; D004847:Epithelial Cells / Q000821:virology*; D053485:Porcine epidemic diarrhea virus / Q000302:isolation & purification*; D012692:Serial Passage; D013552:Swine; D013553:Swine Diseases / Q000821:virology*; D019562:Viral Load; D014776:Virus Cultivation / Q000379:methods*
keywords: isolation and identification; porcine epidemic diarrhea virus (PEDV) NJ strain; porcine intestinal epithelial cells