Development of an ELISA based on the baculovirus-expressed capsid protein of porcine circovirus type 2 as antigen.
The genome of porcine circovirus type 2 (PCV2) contains two major open reading frames, which have been shown to encode the virus capsid and replication-associated proteins. The capsid protein is a major structural protein of the virus; it can be a suitable target antigen for detecting PCV2-specific antibodies to monitor PCV2 infection. To produce the antigen, the capsid protein coding sequence was cloned into a baculovirus transfer vector, and a recombinant capsid (rC) protein of PCV2 was expressed as a combined fusion protein in frame with a C-terminal peptide of six histidines. The affinity-purified rC protein was used as coating antigen to develop an ELISA for detecting the virus-specific antibodies in swine sera. The rC protein-based ELISA (rcELISA) was evaluated by examining a panel of 49 PCV2-positive and 49 PCV2-negative swine sera. In comparative experiments of immunoperoxidase monolayer assay (IPMA) using 102 field sera, there was 89.2% coincidence between data obtained by the rcELISA and IPMA. The rcELISA achieved 88.5% specificity and 89.4% sensitivity for detection of PCV2 antibody in the field sera. The assay showed no cross-reactivity with antibodies to PCV type 1, porcine reproductive and respiratory syndrome virus and porcine parvovirus. The results suggest that the rcELISA is suitable for routine serodiagnosis and epidemiological surveys of PCV2-associated diseases.
Liu
Changming
C
Nippon Institute for Biological Science, Ome, Tokyo, Japan.
Ihara
Takeshi
T
Nunoya
Tetsuo
T
Ueda
Susumu
S
D000818:Animals; D000956:Antigens, Viral / Q000276:immunology*; D016367:Baculoviridae / Q000378:metabolism; D015153:Blotting, Western; D036022:Capsid Proteins / Q000235:genetics*; D017925:Circovirus / Q000235:genetics* / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007124:Immunoenzyme Techniques; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA; D012698:Serologic Tests; D014760:Viral Fusion Proteins / Q000378:metabolism
D000956:Antigens, Viral; D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins