title: A sensitive epitope-blocking ELISA for the detection of Chikungunya virus-specific antibodies in patients.
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.
authors.affiliation: Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia 4072, QLD, Australia.
authors.forename: Lucas Y H
authors.initials: LY
authors.lastname: Goh
authors.affiliation: Laboratory of Microbial Immunity, Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR), Biopolis, Singapore 138648, Singapore.
authors.forename: Yiu-Wing
authors.initials: YW
authors.lastname: Kam
authors.affiliation: Laboratory of Virology, Wageningen University, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.
authors.forename: Stefan W
authors.initials: SW
authors.lastname: Metz
authors.affiliation: Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia 4072, QLD, Australia.
authors.forename: Jody
authors.initials: J
authors.lastname: Hobson-Peters
authors.affiliation: Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia 4072, QLD, Australia.
authors.forename: Natalie A
authors.initials: NA
authors.lastname: Prow
authors.affiliation: School of Pathology and Laboratory Medicine, The University of Western Australia, Crawley 6009, WA, Australia; Division of Microbiology and Infectious Diseases, PathWest Laboratory Medicine, Nedlands 6009, WA, Australia.
authors.forename: Suzi
authors.initials: S
authors.lastname: McCarthy
authors.affiliation: School of Pathology and Laboratory Medicine, The University of Western Australia, Crawley 6009, WA, Australia; Division of Microbiology and Infectious Diseases, PathWest Laboratory Medicine, Nedlands 6009, WA, Australia.
authors.forename: David W
authors.initials: DW
authors.lastname: Smith
authors.affiliation: Laboratory of Virology, Wageningen University, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.
authors.forename: Gorben P
authors.initials: GP
authors.lastname: Pijlman
authors.affiliation: Laboratory of Microbial Immunity, Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR), Biopolis, Singapore 138648, Singapore.
authors.forename: Lisa F P
authors.initials: LF
authors.lastname: Ng
authors.affiliation: Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia 4072, QLD, Australia. Electronic address: roy.hall@uq.edu.au.
authors.forename: Roy A
authors.initials: RA
authors.lastname: Hall
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology*; D065632:Chikungunya Fever / Q000175:diagnosis*; D002646:Chikungunya virus / Q000276:immunology* / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000276:immunology*; D006801:Humans; D012680:Sensitivity and Specificity; D022701:Virosomes / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D022701:Virosomes
keywords: Chikungunya virus; Clinical diagnostic; Epitope-blocking ELISA; Monoclonal antibodies; Virus-like particles