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Journal Articles Viruses Year : 2021

Development and Validation of an ELISA for the Detection of Bluetongue Virus Serotype 4-Specific Antibodies

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Emmanuel Bréard
  • Function : Correspondent author
Georges Beaud
  • Function : Author
Lydie Postic
  • Function : Author
Aurore Fablet
  • Function : Author
Grégory Caignard
Cyril Viarouge
  • Function : Author
Bernd Hoffmann
Damien Vitour
Stéphan Zientara

Abstract

In this article, we describe the development and evaluation of a double antigen sandwich enzyme-linked immunosorbent assay (ELISA) able to detect serotype 4-specific antibodies from BTV-4 infected or vaccinated animals using a recombinant BTV-4 VP2 protein. The coding sequence of VP2 was inserted into a pVote plasmid by recombination in the Gateway® cloning system. Vaccinia virus (VacV) was used as a vector for the expression of the recombinant VP2. After production in BSR cells, recombinant VP2 was purified by immunoprecipitation using a FLAG tag and then used both as the coated ELISA antigen and as the HRP-tagged conjugate. The performance of the ELISA was evaluated with 1186 samples collected from BTV negative, infected or vaccinated animals. The specificity and sensitivity of the BTV-4 ELISA were above the expected standards for the detection of anti-BTV-4 VP2 antibodies in animals reared in Europe or in the Mediterranean basin. Cross-reactions were observed with reference sera for serotypes 10 and 20, and to a lesser extent with serotypes 12, 17 and 24, due to their genetic proximity to serotype 4. Nevertheless, these serotypes have never been detected in Europe and the Mediterranean area. This ELISA, which requires only the production of a recombinant protein, can be used to detect BTV serotype 4-specific antibodies and is therefore an attractive alternative diagnostic method to serum neutralization.

Dates and versions

anses-03897957 , version 1 (14-12-2022)

Licence

Attribution - CC BY 4.0

Identifiers

Cite

Emmanuel Bréard, Mathilde Turpaud, Georges Beaud, Lydie Postic, Aurore Fablet, et al.. Development and Validation of an ELISA for the Detection of Bluetongue Virus Serotype 4-Specific Antibodies. Viruses, 2021, 13 (9), pp.1741. ⟨10.3390/v13091741⟩. ⟨anses-03897957⟩
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