Premium
Development of a Double‐Antigen Microsphere Immunoassay for Simultaneous Group and Serotype Detection of Bluetongue Virus Antibodies
Author(s) -
Breard E.,
Garnier A.,
Despres P.,
Blaise Boisseau S.,
Comtet L.,
Viarouge C.,
BakkaliKassimi L.,
Pourquier P.,
Hudelet P.,
Vitour D.,
Rossi S.,
Belbis G.,
Sailleau C.,
Zientara S.
Publication year - 2017
Publication title -
transboundary and emerging diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.392
H-Index - 63
eISSN - 1865-1682
pISSN - 1865-1674
DOI - 10.1111/tbed.12578
Subject(s) - serology , virology , serotype , immunoassay , antibody , biology , multiplex , antigen , orbivirus , virus , recombinant dna , biotinylation , reoviridae , microbiology and biotechnology , immunology , rotavirus , bioinformatics , biochemistry , gene
Summary Bluetongue viruses ( BTV ) are arboviruses responsible for infections in ruminants. The confirmation of BTV infections is based on rapid serological tests such as enzyme‐linked immunosorbent assays ( ELISA s) using the BTV viral protein 7 ( VP 7) as antigen. The determination of the BTV serotype by serological analyses could be only performed by neutralization tests ( VNT ) which are time‐consuming and require BSL 3 facilities. VP 2 protein is considered the major serotype‐defining protein of BTV . To improve the serological characterization of BTV infections, the recombinant VP 7 and BTV serotype 8 ( BTV ‐8) VP 2 were synthesized using insect cells expression system. The purified antigens were covalently bound to fluorescent beads and then assayed with 822 characterized ruminant sera from BTV vaccinations or infections in a duplex microsphere immunoassay ( MIA ). The revelation step of this serological duplex assay was performed with biotinylated antigens instead of antispecies conjugates to use it on different ruminant species. The results demonstrated that MIA detected the anti‐ VP 7 antibodies with a high specificity as well as a competitive ELISA approved for BTV diagnosis, with a better efficiency for the early detection of the anti‐ VP 7 antibodies. The VP 2 MIA results showed that this technology is also an alternative to VNT for BTV diagnosis. Comparisons between the VP 2 MIA and VNT results showed that VNT detects the anti‐ VP 2 antibodies in an early stage and that the VP 2 MIA is as specific as VNT . This novel immunoassay provides a platform for developing multiplex assays, in which the presence of antibodies against multiple BTV serotypes can be detected simultaneously.