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Expression of virus proteins for ELISA kit development

by | Aug 19, 2021 | Viticulture

Project number
Pathsol 20-01

Project leader
Pietersen, I

Team members

Pietersen, G

Julies, J

le Cordeur, N.S

Truter, T


Completion date


Objectives and Rationale
Viral diseases drastically reduce yields, quality, and longevity in grapevines. Widespread testing is essential for disease control, yet there are currently no local ELISA kits available for testing Grapevine virus A, B, and Fleck. Our aim was to develop ELISA assays capable of detecting these Vitiviruses at least as well as current commercial kits.

Representative viral coat (CP) and movement proteins (MP) of GVA, GVB, and GFkV were expressed in E. coli as antigens, and used to raise antisera in goats and rabbits. These antisera were analysed against known positive and negative controls, and evaluated in parallel with commercial assays to determine their performances.

Key Results
Antisera were successfully raised against several Vitivirus proteins. Low levels of detection were shown for some viral variants/infected grapevines, yet none of the antisera were able to reliably detect their target pathogens. However, GFkV and other antisera provided by Dariusz Goszczynski showed some promising results which will be further explored.

Key Conclusion and Discussion
In spite of strong antibody responses against their respective protein antigens, most of the polyclonal antibodies generated in this project were unable to reliably detect their target Vitiviruses or outperform current commercial kits. However, the performance of some may be enhanced following further optimisations, which will be performed in coming months.

Take Home Message for Industry
Performances of ELISAs developed in this project are currently inadequate to replace commercial kits, but some findings and products generated in this project hold potential for future developments and will be further explored.

Final Report.pdf

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