Development of a Double-Antigen Microsphere Immunoassay for Simultaneous Group and Serotype Detection of Bluetongue Virus Antibodies - Anses - Agence nationale de sécurité sanitaire de l’alimentation, de l’environnement et du travail Accéder directement au contenu
Article Dans Une Revue Transboundary and emerging diseases Année : 2016

Development of a Double-Antigen Microsphere Immunoassay for Simultaneous Group and Serotype Detection of Bluetongue Virus Antibodies

Emmanuel Breard
Aline Garnier
  • Fonction : Auteur
Sandra Blaise-Boisseau
L. Comtet
  • Fonction : Auteur
Cyril Viarouge
  • Fonction : Auteur
Labib Bakkali-Kassimi
P. Pourquier
  • Fonction : Auteur
S. Hudelet
  • Fonction : Auteur
Damien Vitour
  • Fonction : Auteur
G. Belbis
Stéphan Zientara

Résumé

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 (ELISAs) using the BTV viral protein 7 (VP7) 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 BSL3 facilities. VP2 protein is considered the major serotype-defining protein of BTV. To improve the serological characterization of BTV infections, the recombinant VP7 and BTV serotype 8 (BTV-8) VP2 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-VP7 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-VP7 antibodies. The VP2 MIA results showed that this technology is also an alternative to VNT for BTV diagnosis. Comparisons between the VP2 MIA and VNT results showed that VNT detects the anti-VP2 antibodies in an early stage and that the VP2 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.

Dates et versions

anses-01643780 , version 1 (21-11-2017)

Identifiants

Citer

Emmanuel Breard, Aline Garnier, Philippe Desprès, Sandra Blaise-Boisseau, L. Comtet, et al.. Development of a Double-Antigen Microsphere Immunoassay for Simultaneous Group and Serotype Detection of Bluetongue Virus Antibodies. Transboundary and emerging diseases, 2016, 64 (6), pp.1837 - 1847. ⟨10.1111/tbed.12578⟩. ⟨anses-01643780⟩
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