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Expression and diagnostic application of nucleocapsid protein of porcine reproductive and respiratory syndrome virus  

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)
Publication Information
Korean Journal of Veterinary Research / v.43, no.1, 2003 , pp. 129-137 More about this Journal
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
PRRS; nucleocapsid protein; E. coli expression; indirect sandwich enzyme-linked immunosorbent assay;
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