Targeted Design of DIVA Vaccines Using Reverse Genetics

This project involves research to provide the tools needed for the establishment of genetically modified pestiviruses engineered specifically for the requirements of the Differentiate Infected from Vaccinated Animals (DIVA) principle. New innovative strategies are needed to facilitate construction of stable infectious pestivirus clones in order to obtain genetically modified pestiviruses from a wider range of pestiviral strains. The novel full-genome amplification strategy for pestiviruses, that our group has developed recently opens new ways for targeted design of chimeric pestiviruses specifically tailored for use as DIVA vaccines against classical swine fever (CSF). In this project, this novel generic strategy for amplification of pestiviruses will be combined with the innovative BAC (bacterial artificial chromosome) technology. This allows construction of stable infectious clones of large RNA viruses and facilitates specific genetic manipulation hence a new set of molecular tools can be established, which will give increased flexibility in design of new modified pestiviruses for the future generation of DIVA vaccines. Using this strategy, for targeted design of genetically modified pestiviruses, the work can be expedited and focused in principal on any pestiviral strain and hence is not limited to the availability of an existing infectious clone. The long RT-PCR strategy will significantly simplify and streamline the workflow and pave the way for in vitro characterisation and in vivo testing of new and improved DIVA vaccine candidates.