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Successive Passage In Vitro Led to Lower Virulence and Higher Titer of A Variant Porcine Epidemic Diarrhea Virus.
Zhao, Pengwei; Wang, Song; Chen, Zhi; Yu, Jiang; Tang, Rongzhi; Qiu, Wenbin; Zhao, Lu; Liu, Yueyue; Guo, Xiaozhen; He, Hongbin; Xu, Guanlong; Li, Jinxiang; Wu, Jiaqiang.
  • Zhao P; Department of Biochemistry and Molecular Biology, School of Medicine, Zhejiang University, Hangzhou 310058, China.
  • Wang S; Ruminant Diseases Research Center, Key Laboratory of Animal Resistant Biology of Shandong, College of Life Sciences, Shandong Normal University, Jinan 250014, China.
  • Chen Z; Shandong Provincial Key Laboratory of Animal Diseases Control and Breeding, Shandong Academy of Agricultural Sciences, Jinan 250100, China.
  • Yu J; Shandong Provincial Key Laboratory of Animal Diseases Control and Breeding, Shandong Academy of Agricultural Sciences, Jinan 250100, China.
  • Tang R; Shandong Provincial Key Laboratory of Animal Diseases Control and Breeding, Shandong Academy of Agricultural Sciences, Jinan 250100, China.
  • Qiu W; Ruminant Diseases Research Center, Key Laboratory of Animal Resistant Biology of Shandong, College of Life Sciences, Shandong Normal University, Jinan 250014, China.
  • Zhao L; Shandong Provincial Key Laboratory of Animal Diseases Control and Breeding, Shandong Academy of Agricultural Sciences, Jinan 250100, China.
  • Liu Y; Shandong Key Laboratory of Poultry Diseases Diagnosis and Immunology, Shandong Academy of Agricultural Sciences, Jinan 250023, China.
  • Guo X; Shandong Key Laboratory of Poultry Diseases Diagnosis and Immunology, Shandong Academy of Agricultural Sciences, Jinan 250023, China.
  • He H; Shandong Key Laboratory of Poultry Diseases Diagnosis and Immunology, Shandong Academy of Agricultural Sciences, Jinan 250023, China.
  • Xu G; Ruminant Diseases Research Center, Key Laboratory of Animal Resistant Biology of Shandong, College of Life Sciences, Shandong Normal University, Jinan 250014, China.
  • Li J; China Institute of Veterinary Drug Control, Beijing 100081, China.
  • Wu J; National Agricultural Science and Technology Center, Chengdu 610000, China.
Viruses ; 12(4)2020 04 01.
Article in English | MEDLINE | ID: covidwho-833482
ABSTRACT
A highly virulent porcine epidemic diarrhea virus (PEDV) appeared in China and spread rapidly to neighbor countries, which have led to great economic losses to the pig industry. In the present study, we isolated a PEDV using Vero cells and serially propagated 100 passages. PEDV SDSX16 was characterized in vitro and in vivo. The viral titers increased to 107.6 TCID50/mL (100th) by serial passages. The spike (S) gene and the whole gene of the SDSX16 virus was fully sequenced to assess the genetic stability and relatedness to previously identified PEDV. Along with successive passage in vitro, there were 18 nucleotides (nt) deletion occurred in the spike (S) gene resulting in a deletion of six amino acids when the SDSX16 strain was passaged to the 64th generation, and this deletion was stable until the P100. However, the ORF1a/b, M, N, E, and ORF3 genes had only a few point mutations in amino acids and no deletions. According to growth kinetics experiments, the SDSX16 deletion strain significantly enhanced its replication in Vero cells since it was passaged to the 64th generation. The animal studies showed that PEDV SDSX16-P10 caused more severe diarrhea and vomiting, fecal shedding, and acute atrophic enteritis than SDSX16-P75, indicating that SDSX16-P10 is enteropathogenic in the natural host, and the pathogenicity of SDSX16 decreased with successive passage in vitro. However, SDSX16-P10 was found to cause lower levels of cytokine expression than SDSX16-P75 using real-time PCR and flow cytometry, such as IL1ß, IL6, IFN-ß, TNF-α, indicating that SDSX16-P10 might inhibit the expression of cytokines. Our data indicated that successive passage in vitro resulted in virulent attenuation in vivo of the PEDV variant strain SDSX16.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Swine Diseases / Coronavirus Infections / Viral Load / Porcine epidemic diarrhea virus Type of study: Prognostic study / Randomized controlled trials Topics: Variants Limits: Animals Language: English Year: 2020 Document Type: Article Affiliation country: V12040391

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Swine Diseases / Coronavirus Infections / Viral Load / Porcine epidemic diarrhea virus Type of study: Prognostic study / Randomized controlled trials Topics: Variants Limits: Animals Language: English Year: 2020 Document Type: Article Affiliation country: V12040391