The aim of this project was to develop and evaluate diagnostic reagents, tools and methodologies to identify both generic and specific strains of PRRSV as well as future proofing this technology for the rapid identification of new and emerging strains. The plan of work and objectives consisted of expressing different PRRSV proteins which might be used to detect strain specific and non-specific variants. The candidate proteins selected were the virus genome binding protein (nucleocapsid (N) protein) and surface proteins of the virus particle (e.g. GP5 protein). Apart from traditional expression technologies the focus of this research was to pioneer the use of recently developed rapid in vitro (i.e. test tube) protein expression systems to demonstrate proof of principal in identifying new strains of PRRSV. Traditional virus protein expression technologies were used to produce differentially modified proteins (as occurs in the infected cell) for use in ELISA and to act as comparison for proteins produced in the rapid expression systems. The data indicated that whilst protein modification resulted in increases in efficiency for ELISA detection, the rapid protein expression system was by far the most efficient way of producing protein quickly. Indeed from receipt of viral gene sequence purified protein suitable for ELISA was produced within 5 days in the rapid system, compared to approximately up to a month for lead in times for conventional protein expression. As highlighted in our publication describing rapid protein expression, this technology thus holds great promise for responding rapidly to detect new and emerging strains of PRRSV. Future work highlighted in the grant and planned for these PRRSV proteins was to characterize them at the cell biological level in order to better understand their roles in the life cycle of PRRSV and for exploiting these biological properties for vaccine development. This work was conducting in collaboration with the bio-imaging unit at the University and was the first to characterize the real time trafficking of N protein around the cell. As highlighted our the publication of this work, this information can be rapidly exploited for the development of growth attenuated recombinant vaccines. Contact: Dr. Julian A. Hiscox, University of Leeds, [email protected].
Contribute to Pork Research
Discover how you can help improve the pork industry by checking out research RFPs or sharing your own ideas. If you have a research idea, we want to know! Here’s your chance to make a difference.