Universal method of purification and concentration of viruses on the example of innovation for geese parvovirus

Abstract

A new method of purification and concentration of goose enteritis virus has been developed. For this purpose, three options for further purification of the virus were used and their comparative analysis was performed. The virus was purified without detergents, as well as with detergents – sarcosyl and nonide R-40. We obtained the best results using the mild nonionic detergent nonidet R-40, which was used in our further work. The virus was identified by electrophoretic studies in polyacrylamide gel, as well as electron microscopy. During purification and concentration of the virus, the infectious titer of the virus was 9.2-9.5 lg TCD50/cm3 suspension, in which the cell monolayer in 50% of the wells was affected by cytopathic action), which is 2 lg TCD50/cm3 higher than in the original material. The protein content in the test samples ranged from 200 to 500 mg/ ml. Thus, analyzing our results on the purification and concentration of goose enteritis virus, we can conclude that the antigen obtained by this method is suitable for the development of enzyme-linked immunosorbent assay (ELISA). Hyperimmune and negative sera for ELISA diagnostics were obtained, the optimal ratios of components for designing an ELISA test system were worked out, and the formula for recalculating antibody titers in geese blood sera when testing them in one dilution was derived. A positive-negative threshold was determined for this diagnosticum (which of the studied sera have a positive, doubtful or negative titer of antibodies to the causative agent of viral enteritis in geese). In the new conditions the spread of particularly dangerous viruses, this development, with the appropriate equipment, can be further used to purify and concentrate these viruses, study their biological properties, cultivate and use them in the development of new vaccines.