The invention discloses an animal epidemic disease three-color fluorescence RT-PCR detection kit and a detection method thereof, and is characterized in that the kit comprises multiple fluorescence RT-PCR reaction mother liquor, a positive control, a negative control, an AMV reverse transcriptase, and a Taq polymerase, wherein the multiple fluorescence RT-PCR reaction mother liquor contains a multiple 10*fluorescence RT-PCR reaction buffer, an avian influenza primer and a probe, a newcastle disease virus primer and a probe, an avian infectious bronchitis primer and a probe, and bovine serum albumin, wherein sequences of the forward primers, reverse primers, and specific probes are as shown in SEQIDNO. 1, NO. 2, NO.3, NO.4, NO.5, NO.6, NO.7. NO.8, NO.9, and NO.10. The advantages of the invention are that avian influenza, newcastle disease, and avian infectious bronchitis viruses can be detected simultaneously in a same reaction tube; the specificity is strong; the sensitivity is high, is rapid and accurate.