The invention relates to the technical field of microbiological detection of veterinary pathogens and discloses a reagent and method for identifying a rabies virus vaccine strain and a wild type virusstrain. The reagent disclosed by the invention consists of an upstream primer of which the nucleotide sequence is shown as SEQ ID NO:1, a downstream primer of which the nucleotide sequence is shown as SEQ ID NO:2, a rabies virus vaccine strain probe of which the nucleotide sequence is shown as SEQ ID NO:3 and is modified with a fluorescent group 1 and a quenching group, and a rabies virus wild type virus strain of which the nucleotide sequence is shown as SEQ ID NO:4, and is modified with a fluorescent group 2 and a quenching group. The invention provides one group of primers and probe reagents with higher specificity and sensitivity; by means of the reagent, the aims of accurately identifying the rabies virus vaccine strain and the wild type virus strain are achieved, and the defects that the wild virus strain is difficulty distinguished from the vaccine strain by existing general methods such as virus separation, serological tests and an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) method are made up; the reagent and the method can be comprehensively applied to safety evaluation and clinical diagnosis of a rabies virus vaccine.