Single dilution Avidity-Blocking ELISA as an alternative to the Bovine Viral Diarrhea Virus neutralization test

J Virol Methods. 2011 Aug;175(2):228-35. doi: 10.1016/j.jviromet.2011.05.022. Epub 2011 May 19.

Abstract

This study describes the development and validation of a blocking ELISA that measures avidity of BVDV-specific immunoglobulins (Igs) as an alternative to the classic virus neutralization test. The assay comprises a recombinant soluble E2 glycoprotein as target antigen, a neutralizing serum as detector antibody and a washing-step with a chaotropic agent to determine BVDV-specific Igs avidity. Avidity-Blocking ELISA was validated with 100 negative and 87 positive BVDV-neutralization serum samples from either infected or vaccinated bovines (inactivated commercial vaccines). Specificity and sensitivity of the Avidity-Blocking ELISA were 100% and 98.8%, respectively. The assay was standardized to use a single dilution, so that 90 samples can be tested per plate. Results expressed as Avidity Index (AI) correlated with BVDV neutralizing titers (r=0.94). Unlike the virus neutralization test, the Avidity-Blocking ELISA could discriminate between infected and vaccinated animals (DIVA), suggesting that avidity measurement can be a valuable tool to achieve DIVA compliances. The data show that the avidity of anti BVDV antibodies is related to their capacity to block viral infection in vitro.

Publication types

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

MeSH terms

  • Animals
  • Antibodies, Viral / blood*
  • Bovine Virus Diarrhea-Mucosal Disease / diagnosis*
  • Bovine Virus Diarrhea-Mucosal Disease / immunology
  • Bovine Virus Diarrhea-Mucosal Disease / virology
  • Cattle
  • Clinical Laboratory Techniques / methods*
  • Diarrhea / virology*
  • Diarrhea Viruses, Bovine Viral / immunology*
  • Enzyme-Linked Immunosorbent Assay / methods
  • Neutralization Tests / methods
  • Sensitivity and Specificity

Substances

  • Antibodies, Viral