The invention provides a primer combination for detecting the instability of a microsatellite, a kit, a detection method and application of the primer combination, the kit and the detection method. The primer combination comprises primer pairs shown in SEQ ID NO:1-18 for NR21, NR24, NR27, BAT25, BAT26, MONO-27, Penta D, Penta E and Amel. According to the primer combination, primers are designed for six SNP loci of NR21, NR24, NR27, BAT25, BAT26 and MONO-27; according to the states of the six SNP loci, the instability of the microsatellite is evaluated; according to three contrast polymorphismloci of Penta D, Penta E and Amel, the consistency of a to-be-detected tissue sample and a contrast sample thereof is evaluated; the accuracy is high, the number of used samples is low, the amplification efficiency is high, strips are clear, and the sensitivity is high.