The invention discloses a multiplex RT-PCR kit for BTV-11, BTV-17, BTV-20, BTV-23 and BTV-24 genotype identification and a detection method of the multiplex RT-PCR kit. The multiplex RT-PCR kit is used for simultaneously discriminating and detecting bluetongue virus types 11, 17, 20, 23 and 24 with a single tube. According to the method, 5 pairs of PCR specific primers are designed according to conserved regions of VP2 gene sequences of various types of viruses, in addition, a pair of non-biogenic-derivation universal primers are synthesized with reference to a GeXP principle, the universal primers are separately added to upstream and downstream of each pair of specific primers to form 5 pair of specific embedded primers, reverse transcription is carried out with the specific primers, andfinally, multiplex PCR is constructed by adopting the universal primers and the specific embedded primers. By utilizing an optimized multiplex PCR system and conditions, one or more of the 5 genotypesof the bluetongue viruses can be simultaneously discriminated and detected with the single tube, specific amplification to other types of BTV, PPRV and FMDV nucleic acid is avoided, and the minimum detection concentration can reach a pg level. The method disclosed by the invention is high in sensitivity, high in specificity and easy in result observation and is timesaving and laborsaving.