The invention provides a fluorescence PCR detection kit for human K-RAS gene mutation. The kit comprises a forward primer sequence K-M1 PCR reaction solution, a forward primer sequence K-M2 PCR reaction solution, a forward primer sequence K-M3 PCR reaction solution, a forward primer sequence K-M4 PCR reaction solution, a forward primer sequence K-M5 PCR reaction solution, a forward primer sequence K-M6 PCR reaction solution and a forward primer sequence K-M7 PCR reaction solution which are used for detecting Gly12Asp mutation, Gly12Val mutation, Gly12Ser mutation, Gly12Cys mutation, Gly12Ala mutation, Gly12Arg mutation and Gly13Asp mutation on a codon 12 and codon 13. According to the kit, under the 100ng/reaction wild-type human DNA background, amplification signals and the false positive condition are avoided, and it shows that the kit is good in specificity and high in antijamming capability. By the adoption of the kit, human K-RAS gene mutation in samples such as clinical paraffin sections can be detected quickly, and a reliable experiment basis is provided for diagnosing related cancer therapies.