[Isolation and multiplication of rabies virus in cell lines under conditions of routine rabies diagnosis].

Isolation of rabies street virus from brains of animals with rabies was performed on cells of mouse neuroblastoma, infecting cells with suspension of tested brain tissue diluted 1:10, incubating at 37 degrees C in standard incubator (culture in Legroux bottles and Leighton tubes with rubber stoppers) and in 37 degrees C in an incubator with moist chamber and regulated flow of carbon dioxide (Mirco-Well plates, bottom-flat tubes, bottles produced by Nuc, 25-chamber plates produced by Flow). The main conditions for obtaining positive results was proper selection and composition of growing fluid for cells (MEM with Earle's salts produced by Sigma enriched with vitamins, L-glutamine, amino acids and calf fetal fluid) and limited number of passages of tissue after thawing cells from the bank. With passaging neuroblastoma cells are becoming resistant to infection. Growth of cells abd isolation of the virus were possible both in atmosphere containing 5% CO2 and normal air, but application of a standard incubator required higher usage of growth fluids. Positive results were also obtained with infected material stored for a long time. Storage of material at 37 degrees C was making isolation more difficult. Multiplication of stable viruses on BHK, neuroblastoma and Vero cells permitted for obtaining antigens for serological diagnosis of rabies in vitro.