The invention discloses a breast cancer 21 gene detection kit and a detection method thereof. The kit comprises primers and probes for detecting genes ACTB, GAPDH, RPLPO, GUS, TFRC, CD68, BAG1, GSTM1,HER2, GRB7, MMP11, CTSL2, KI67, STK15, CCNB1, MYBL2, Survivin, ESR1, PGR, BCL2 and SCUBE2. Two pairs of primers and two specific probes are respectively designed for 21 genes in the kit, a primer andprobe liquid of each gene is placed in one reaction tube, a same buffer solution and a procedure are adopted for QPCR (Quantitative Polymerase Chain Reaction) amplification, the operation is simple and convenient to carry out, the detection time can be shortened, and clear and accurate results can be achieved; the kit is wide in application, and applicable to both scientific research and clinicaldetection and applicable to single or multiple samples, and rapid and accurate detection results can be achieved.