A sensitive epitope-blocking ELISA for the detection of Chikungunya virus-specific antibodies in patients

J Virol Methods. 2015 Sep 15:222:55-61. doi: 10.1016/j.jviromet.2015.05.011. Epub 2015 May 27.

Abstract

Chikungunya fever (CHIKF) has re-emerged as an arboviral disease that mimics clinical symptoms of other diseases such as dengue, malaria, as well as other alphavirus-related illnesses leading to problems with definitive diagnosis of the infection. Herein we describe the development and evaluation of a sensitive epitope-blocking ELISA (EB-ELISA) capable of specifically detecting anti-chikungunya virus (CHIKV) antibodies in clinical samples. The assay uses a monoclonal antibody (mAb) that binds an epitope on the E2 protein of CHIKV and does not exhibit cross-reactivity to other related alphaviruses. We also demonstrated the use of recombinant CHIK virus-like particles (VLPs) as a safe alternative antigen to infectious virions in the assay. Based on testing of 60 serum samples from patients in the acute or convalescent phase of CHIKV infection, the EB-ELISA provided us with 100% sensitivity, and exhibited 98.5% specificity when Ross River virus (RRV)- or Barmah Forest virus (BFV)-immune serum samples were included. This assay meets the public health demands of a rapid, robust, sensitive and specific, yet simple assay for specifically diagnosing CHIK-infections in humans.

Keywords: Chikungunya virus; Clinical diagnostic; Epitope-blocking ELISA; Monoclonal antibodies; Virus-like particles.

Publication types

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

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Antibodies, Viral / blood*
  • Antibodies, Viral / immunology
  • Antigens, Viral / immunology*
  • Chikungunya Fever / diagnosis*
  • Chikungunya virus / immunology*
  • Chikungunya virus / isolation & purification
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Epitopes / immunology*
  • Humans
  • Sensitivity and Specificity
  • Virosomes / genetics
  • Virosomes / immunology

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Antigens, Viral
  • Epitopes
  • Virosomes