›› 2019, Vol. 46 ›› Issue (2): 582-589.doi: 10.16431/j.cnki.1671-7236.2019.02.030

• Preventive Veterinary Medicine • Previous Articles     Next Articles

Isolation and Identification of Porcine Epidemic Diarrhea Virus Strain JS2017 and Its S Gene Sequence Analysis

ZHU Haixia   

  1. Fuzhou Dabeinong Biotechnology Co., Ltd., Fuzhou 350014, China
  • Received:2018-08-24 Online:2019-02-20 Published:2019-02-20

Abstract:

In order to understand the genetic variation of porcine epidemic diarrhea virus (PEDV) in a large-scale pig farm in Jiangsu,the intestinal content of piglets with watery diarrhea were collected and detected for PEDV by RT-PCR.The diseased tissue detected as positive was treated with 20 μg/mL trypsin,and then inoculated into Vero cells for culture.After the cell culture was repeatedly frozen and thawed,the virus solution was harvested and identified by RT-PCR.The S gene was amplified by a segmentation overlap strategy,and the nucleotide homology and genetic evolution of S gene was analyzed.The results showed that the pathological tissue was positive for PEDV by RT-PCR.After 4 generations of inoculation with Vero cells,the positive pathogens showed obvious cytopathic changes,which showed cell syncytosis,vacuolization and finally lysis.After the harvested virus solution was identified by RT-PCR,it was confirmed that one PEDV was isolated and designated as JS2017 strain.Sequence analysis showed that the S gene sequence of JS2017 strain was in the same evolutionary branch as the American variant strain OH1414,Canadian variant strain ON-007,Chinese variant strain PEDV-LYG and CH/PDS/2015,and its nucleotide homology was as high as 99.0%.Among them,the Chinese mutant strain YC2014 had the closest relationship,and its nucleotide homology was 100.0%;JS2017 strain had low homology with the classical strains CV777 and DR13,and had a distant relationship.The results showed that the JS2017 strain isolated in this study was a PEDV local epidemic strain,which was closely related to the 2014 Jiangsu isolate (YC2014 strain), and had a far-reaching relationship with the current swine epidemic diarrhea vaccine strain.

Key words: porcine epidemic diarrhea virus; S gene; variant strain

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