Development and evaluation of a VP1-ELISA for detection of antibodies to duck hepatitis type 1 virus

J Virol Methods. 2010 Oct;169(1):66-9. doi: 10.1016/j.jviromet.2010.06.018. Epub 2010 Jul 4.

Abstract

The VP1-encoding gene of the duck hepatitis type 1 virus (DHV-1) HP-1 strain was cloned and expressed in Escherichiacoli. The open reading frame (ORF) of VP1 comprised 714 bp and encoded 238 amino acids, with a predicated molecular mass of 26.5 kDa. The expressed VP1 fusion protein in E. coli was detected by Western blotting with duck anti-DHV-1 polyclonal serum. A VP1-ELISA using the expressed VP1 protein as a coating antigen for the detection of antibodies to DHV-1 in ducks was developed. In comparison with the virus neutralization test, the specificity and sensitivity of the VP1-ELISA was 92.5% and 96.7%. Comparative analysis between Western blots and the VP1-ELISA showed that the concordance between the two methods was 86%. The VP1-ELISA did not react with the anti-sera to other duck viral pathogens, implying that this protein is specific for the recognition of duck anti-DHV-1 antibodies. Taken together, the VP1-ELISA is a highly sensitive and specific test that could be used for screening for DHV-1 infection and monitoring antibody titres against DHV-1.

Publication types

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

MeSH terms

  • Animals
  • Antibodies, Viral / blood*
  • Antigens, Viral* / chemistry
  • Antigens, Viral* / genetics
  • Antigens, Viral* / immunology
  • Capsid Proteins* / chemistry
  • Capsid Proteins* / genetics
  • Capsid Proteins* / immunology
  • Cloning, Molecular
  • Ducks
  • Enzyme-Linked Immunosorbent Assay / methods
  • Escherichia coli / genetics
  • Gene Expression
  • Hepatitis Virus, Duck / immunology*
  • Hepatitis Virus, Duck / isolation & purification*
  • Molecular Weight
  • Neutralization Tests
  • Open Reading Frames
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Sensitivity and Specificity
  • Virology / methods*

Substances

  • Antibodies, Viral
  • Antigens, Viral
  • Capsid Proteins
  • Recombinant Proteins