Process for propagating rabies virus

摘要

Culture liquids containing live rabies virus are produced by incubating an embryonic cell monolayer infected with live rabies virus in a sterile, protein-free, aqueous, cell-maintenance medium containing at least 0.3% by weight of pancreatic digest of casein under aseptic conditions until a substantial quantity of live rabies virus is present in the culture liquid and removing the culture liquid containing the live rabies virus from the infected, intact, embryonic cell monolayer without disrupting the monolayer. Incubation is at 30 DEG to 37 DEG C., and preferably 32 DEG C., for 1 to 10 and preferably 1 to 7 days. Further maintenance medium is added to the intact, embryonic cell monolayer to replace the removed culture liquid, the culture is incubated until a substantial quantity of live rabies virus is present in the culture liquid and the culture liquid is again removed from the infected, intact, embryonic cell monolayer. The process is repeated until the cell monolayer degenerates. The culture liquid, or a concentrate thereof, may be converted to a killed rabies virus vaccine in a conventional manner, e.g. by using ultra-violet radiation. The embryonic cell monolayers are prepared by cultivating cells obtained from embryonic tissue in an aqueous cell-maintenance medium containing serum in known manner. Preferred embryos are ovian, particularly the embryos of chickens and ducks. The preferred inoculum is C.V.S. fixed rabies virus seed (20% mouse brain suspension containing 2% horse serum). The cell-maintenance medium may be medium No. 199, Eagle's medium, Melrick's SM-2 medium and Hank's Balanced Salt solution. Antibiotics such as penicillin and streptomycin may be present to minimize bacterial contamination.