Development and application of a double-antigen sandwich enzyme-linked immunosorbent assay for detection of antibodies to porcine circovirus 2

Clin Vaccine Immunol. 2012 Sep;19(9):1480-6. doi: 10.1128/CVI.00234-12. Epub 2012 Jul 18.

Abstract

A double-antigen sandwich enzyme-linked immunosorbent assay (ELISA) is described for detection of porcine circovirus 2 (PCV2) antibodies using the well-characterized recombinant PCV2 capsid protein. In a comparative test of 394 pig sera against an indirect immunofluorescence (IIF) test and a commercial ELISA kit (also based on the recombinant PCV2 capsid protein), the results showed that the diagnostic sensitivity, specificity, and accuracy of the assay were, respectively, 90.61, 94.02, and 91.62% compared with IIF and 94.38, 95.28, and 94.67% compared with the commercial ELISA kit. Assay of 12 PCV-free pigs over a 5-week period produced only PCV2-negative titers by all 3 methods. These results and the seroprofiles of 4 pig farms obtained by both the commercial ELISA kit and the double-antigen sandwich ELISA indicate that the sandwich ELISA is a reliable method for detection of antibodies to PCV2. Additionally, the method described here permits the use of undiluted test serum samples simultaneously loaded with horseradish peroxidase (HRP)-conjugated antigen into the test well, and the complete test procedure can be performed in less than 90 min. This double-antigen sandwich ELISA should be a useful tool to aid swine industry professionals in deciding the intervention strategies for the control of PCV2-associated diseases.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Viral / blood*
  • Capsid Proteins* / genetics
  • Circoviridae Infections / diagnosis
  • Circoviridae Infections / veterinary*
  • Circovirus / immunology*
  • Enzyme-Linked Immunosorbent Assay / methods
  • Recombinant Proteins / genetics
  • Sensitivity and Specificity
  • Swine
  • Swine Diseases / diagnosis*
  • Swine Diseases / virology
  • Veterinary Medicine / methods*

Substances

  • Antibodies, Viral
  • Capsid Proteins
  • Recombinant Proteins