Serodiagnostic comparison of enzyme-linked immunosorbent assay and surface plasmon resonance for the detection of antibody to porcine circovirus type 2.
This paper describes the cloning and expression of the capsid protein of Porcine circovirus type 2 (PCV2) in an Escherichia coli expression system that was used to produce a fusion protein for subsequent immunologic studies: enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR). Polymerase chain reaction was used to amplify the gene encoding the capsid protein from the DNA of PCV2. The protein was then cloned into a pRSET prokaryotic expression vector. Western blot analysis revealed that the recombinant protein gave strong signals on a polyvinylidene difluoride membrane when exposed to the serum from a pig infected with PCV2. The expressed protein was purified and used as an antigen for the ELISA and SPR study. A protein chip based on SPR was developed, and the diagnostic potential of SPR was compared with that of ELISA with the use of 70 serum samples obtained from 6 pig farms. There was a strong positive correlation between the ELISA and SPR titers (r = 0.877, P < 0.01). Therefore, this recombinant capsid protein can be used as an antigen for serologic studies, and the SPR, a label-free method, appears to be a valuable and reproducible tool in the serodiagnosis of a PCV2 infection.
Cho
Ho-Seong
HS
College of Veterinary Medicine, Chonnam National University, Gwangju.
Kim
Tae-Jung
TJ
Lee
Jae-Il
JI
Park
Nam-Yong
NY
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D036022:Capsid Proteins / Q000235:genetics; D018173:Circoviridae Infections / Q000175:diagnosis / Q000453:epidemiology / Q000662:veterinary*; D017925:Circovirus / Q000235:genetics / Q000276:immunology*; D004279:DNA, Viral / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005260:Female; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D016036:Seroepidemiologic Studies; D020349:Surface Plasmon Resonance / Q000379:methods / Q000662:veterinary*; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000453:epidemiology*
D000914:Antibodies, Viral; D036022:Capsid Proteins; D004279:DNA, Viral