The invention provides a multiplex PCR combined pyrosequencing kit used for guiding individualized medication of warfarin and a detection method thereof, relating to warfarin. The kit comprises a whole blood genome DNA extraction reagent, multiplex PCR amplification primers, a multiplex PCR amplification reagent, a single-stranded DNA separating and purifying reagent, pyrosequencing primers, a pyrosequencing reagent and a kit body. The detection method comprises the following steps: extraction of human whole blood genome DNA; multiplex PCR amplification; separation and purification of a single-stranded DNA sample; and pyrosequencing and result analysis. According to the invention, main factors influencing difference of individual dosage of warfarin, i.e., important gene polymorphic sites of VKORC1 and CYP2C9, specifically being VKORC1-1639G>A, CYP2C9*2 and CYP2C9*3, are detected so as to guide individualized dosage of warfarin in treatment of patients needing to take warfarin, thereby achieving the purpose of reduction in side effects.