A novel monoclonal antibodies-based sandwich ELISA for detection of serotype 4 fowl adenovirus

Avian Pathol. 2019 Jun;48(3):204-208. doi: 10.1080/03079457.2019.1566595. Epub 2019 Feb 4.

Abstract

As a major causative agent for hepatitis-hydropericardium syndrome (HPS) in chickens, serotype 4 fowl adenovirus (FAdV-4) has caused huge economic losses in the poultry industry globally. However, there is no commercial diagnostic test for FAdV-4 antigens. To generate a rapid approach for specific detection of FAdV-4, a monoclonal antibodies (mAbs)-based sandwich ELISA was developed. In this ELISA, a purified mAb 4A3 and a HRP-labelled mAb 3C2 specific to the fiber-2 of FAdV-4 were used as the capture antibody and detection antibody respectively. Specificity assay revealed the ELISA only reacted with FAdV-4, not with other avian viruses tested. Sensitivity assay showed the limit of detection of the ELISA was 1000 TCID50/ml and 12.5 ng/ml for the FAdV-4 and the purified GST-Fiber2 protein respectively. Moreover, the ELISA could be efficiently applied in detecting the FAdV-4 in tissue samples from a clinically-diseased chicken flock. All these data demonstrated that the ELISA developed here provides a promising tool for rapid and efficient diagnosis of clinical infection with FAdV-4.

Keywords: Serotype 4 fowl adenovirus; monoclonal antibody; sandwich ELISA.

MeSH terms

  • Adenoviridae Infections / diagnosis
  • Adenoviridae Infections / veterinary*
  • Adenoviridae Infections / virology
  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antibodies, Viral / immunology*
  • Aviadenovirus / genetics
  • Aviadenovirus / immunology*
  • Aviadenovirus / isolation & purification
  • Chickens / virology*
  • Enzyme-Linked Immunosorbent Assay / veterinary*
  • Poultry Diseases / diagnosis*
  • Poultry Diseases / virology
  • Sensitivity and Specificity
  • Serogroup

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral