›› 2016, Vol. 43 ›› Issue (2): 356-362.doi: 10.16431/j.cnki.1671-7236.2016.02.010

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Cloning and Sequence Analysis of ORF3 and M Gene of Porcine Epidemic Diarrhea Virus in Guizhou Province

FENG Xu-fang1,2, ZHOU Bi-jun1,2, ZHANG Shuang-xiang3, WANG Yue-zhang4, WEN Ming1,2, CHENG Zhen-tao1,2, MAO Li-hong5, WANG Kai-gong1,2   

  1. 1. College of Animal Science, Guizhou University, Guiyang 550025, China;
    2. Guizhou Pig Quality Safety Engineering Research Center, Guiyang 550025, China;
    3. Animal Epidemic Disease Control Center of Guizhou Province, Guiyang 550001, China;
    4. Qian Chang Stockbreeding Develop Limited Liability Company of Duyun, Duyun 558000, China;
    5. Agricultural Bureau of Huaxi, Guiyang 550025, China
  • Received:2015-07-28 Online:2016-02-20 Published:2016-03-02

Abstract: To study the genetic variations of porcine epidemic diarrhea virus (PEDV) ORF3 and M gene in Guizhou province,we used RT-PCR method to detect PEDV in the dung what collected from diarheal porket in five regions of Guizhou province between April 2014 to March 2015,then selected eight positive samples,cloned and sequenced their ORF3 and M gene.The results showed that 75 samples were positive for PEDV,and the positive rate was 71.43%.The result of sequencing showed that ORF3 and M gene were intact;ORF3 gene shared from 95.1% to 100.0% nucleotide identity and 95.1% to 99.6% amino acid identity,and M gene shared from 98.4% to 100.0% nucleotide identity and 98.7% to 100.0% amino acid identity with eight PEDV Guizhou strains.Phylogenetic analysis revealed that Guizhou strains seem to be closely related to Chinese strains,Korean strains and Thai strains,and there were genetically different from the vaccine strains attenuated DR13 and CV777.The results suggested that in rencent years the mainly etiology of orket diarrhea was velogenic PEDV.

Key words: porcine epidemic diarrhea virus; ORF3 gene; M gene; cloning; sequence analysis

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