Development and evaluation of an in vitro virus isolation procedure as a replacement for the mouse inoculation test...

A tissue culture virus isolation procedure for rabies street strain virus on mouse neuroblastoma cells is described. Parameters for the optimum sensitivity of the procedure were determined to include a minimum 4-day incubation of virus in tissue culture and the use of diethylaminoethyl-dextran for increased cell susceptibility. The in vitro procedure performed well in a comparison with the fluorescent-antibody test and the mouse inoculation test (MIT) on weakly positive brain tissue. Decomposed specimens and virus inhibitors present in brain suspensions were found to interfere with the in vitro procedure. A Formalin-methanol fixative was found to be superior on plastic 96-well plates to previously used fixatives. A 2-year clinical trial of the procedure in parallel with the MIT demonstrated the practicality of the procedure. Accordingly, the New York State rabies diagnostic laboratory has replaced the MIT with the in vitro procedure as a backup for the fluorescent-antibody test in the routine diagnosis of rabies.