| Expression and diagnostic application of nucleocapsid protein of porcine reproductive and respiratory syndrome virus |
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Hyo-Sun Park1, Tae-Uook Hahn2, Hyun-Soo Kim3, Kang-Seuk Choi4, Eun-Jeong Lee5, Shien-Young Kang1 |
1Research Institute of Veterinary Medicine/College of Veterinary Medicine, Chungbuk National University
2Department of Veterinary Medicine, Kangwon National University
3College of Veterinary Medicine, Chungnam National University
4National Veterinary Research and Quarantine Service
5Chungbuk Veterinary Service Laboratory |
| 돼지 생식기호흡기증후군 바이러스의 Nucleocapsid 단백질 발현 및 진단적 응용 |
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박효선1, 한태욱2, 김현수3, 최강석4, 이은정5, 강신영1 |
1충북대학교 수의과대학/동물의학연구소
2강원대학교 수의학과
3충남대학교 수의과대학
4국립수의과학검역원
5충북가축위생시험소 |
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| Abstract |
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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. |
| Key Words:
PRRS, nucleocapsid protein, E. coli expression, indirect sandwich enzyme-linked immunosorbent assay |
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