The aim of this research was to compare the effectiveness of four methods of laboratory diagnostics of rabies - the mouse intracerebral inoculation test (MICIT, MIT), the rabies tissue culture infection test (RTCIT), the rapid rabies enzyme immune diagnosis test (RREID) and the nested reverse transcription polymerase chain reaction (nRT-PCR). The RREID method was carried out in two variants: detection of the rabies antigen in a clarified (centrifugated) and in a non-clarified (noncentrifugated) brain suspension. Three autochthonous street isolates of rabies virus from red foxes and a lynx were used. There was a relative correlation of the diagnostic effectiveness of the two standard methods, MICIT and RTCIT, with standard MICIT being the more sensitive one, RTCIT having other advantages (among others the speed of performance) and thus being preferred. The sensitivity of the RREID method proved to be rather low. If used with noncentrifugated brain suspensions this method may yield nonspecific reactions. If compared particularly with RREID, the nRT-PCR is characterized by a considerably higher diagnostic effectiveness. The sensitivity of nRT-PCR is not affected by preliminary clarification of the brain suspension.