Korean Journal of Veterinary Research (대한수의학회지)

The Korean Society of Veterinary Science (대한수의학회)
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Expression and diagnostic application of nucleocapsid protein of porcine reproductive and respiratory syndrome virus

돼지 생식기호흡기증후군 바이러스의 Nucleocapsid 단백질 발현 및 진단적 응용

  • Park, Hyo-Sun (Research Institute of Veterinary Medicine/College of Veterinary Medicine, Chungbuk National University) ;
  • Hahn, Tae-Uook (Department of Veterinary Medicine, Kangwon National University) ;
  • Kim, Hyun-Soo (College of Veterinary Medicine, Chungnam National University) ;
  • Choi, Kang-Seuk (National Veterinary Research and Quarantine Service) ;
  • Lee, Eun-Jeong (Chungbuk Veterinary Service Laboratory) ;
  • Kang, Shien-Young (Research Institute of Veterinary Medicine/College of Veterinary Medicine, Chungbuk National University)
  • 박효선 (충북대학교 수의과대학/동물의학연구소) ;
  • 한태욱 (강원대학교 수의학과) ;
  • 김현수 (충남대학교 수의과대학) ;
  • 최강석 (국립수의과학검역원) ;
  • 이은정 (충북가축위생시험소) ;
  • 강신영 (충북대학교 수의과대학/동물의학연구소)
  • Accepted : 2003.03.06
  • Published : 2003.03.31
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Abstract

Porcine reproductive and respiratory syndrome (PRRS) is characterized by reproductive failures in sows and respiratory problems in piglets. The nucleocapsid(N) protein, encoded by the open reading frame 7 (ORF7) gene, is known to be the most abundant and antigenic protein in PRRS virus. Therefore, it was suggested that the N protein could be a suitable candidate for the detection of PRRS virus-specific antibodies and diagnosis of PRRS. In the present study, the ORF7 gene encoding the N protein was cloned and expressed as a fusion protein with the glutathione S-transferase (GST) in Escherichia coli. The resulting GST-N recombinant protein was used as an antigen for an indirect sandwich enzyme-linked immunosorbent assay (i-ELISA). Expressed GST-N recombinant protein was migrated at 41 kDa and reacted with ORF7-specific monoclonal antibody by Western blotting. In order to increase the specificity of the ELISA for the detection of PRRS virus-specific antibodes, an i-ELISA was developed using an anti-GST antibody as a capture antibody. The sensitivity and specificity of developed i-ELISA were 92% and 96%, respectively. Based on these results, it was suggested that the i-ELISA is a simple and rapid test for screening a large number of swine sera for the anti-PRRS virus antibodies.

Keywords

References

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