Evaluation of methods of avian leucosis virus inactivation in production of influenza vaccines

The process of production of inactivated influenza vaccines involves a stage of inactivation of both the influenza virus and the possible viral contaminants that can come from the raw materials (chicken embryos). One of such contaminants is the avian leucosis virus. The minimum viral contaminant load reduction that the inactivating agents should guarantee is by 4 lg/ml; this or higher level of the deactivating ability ensures the finished vaccine is free from viral contaminants. The purpose of this work was to cultivate the leucosis virus to the titer of 5 lg/ml (minimum) and to measure the reduction of the avianleucosis virus titer in influenza vaccine intermediates upon exposure to the inactivating agents. The RAV-1 and RAV-2 leucosis virus strains and influenza vaccine intermediates such as virus[1]containing allantoic fluid and virus concentrates were used in the study. Avian leucosis virus titers were determined by enzyme immunoassay. We created conditions for cultivation of the RAV-1 and RAV-2 avian leucosis virus strains in the primary culture of chicken embryo fibroblasts (CEF); the inactivating agents considered were the most commonly used β-propiolactone and UV radiation. It was found that after 12 hours of exposure to β-propiolactone, the RAV-1 avian leucosis virus load decreased by 4.61 ± 0.46 lg, and that of RAV-2 strain — by 4.33 ± 0.33 lg, which indicates that β-propiolactone is an effective inactivating agent. Five minutes of exposure to UV radiation reduces the RAV-1 strain viral load by 4.22 ± 0.31 lg and RAV-2 strain viral load by 4.44 ± 0.48 lg.

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