Hyper-blocking fluorescence quantitative PCR method with high sensitivity for detecting rare mutation

The invention discloses a hyper-blocking fluorescence quantitative PCR method with high sensitivity for detecting rare mutation. The invention adopts specific primers and a probe technology and can rapidly detect gene point mutation in a blood sample. The method has the advantage that (1) the conventional ARMS primers are modified by LNA or mismatched bases are introduced into 3' tail 2th site totail 3th site so that the sensitivity of the mutation detection is not influenced and the specificity of the single base mutation based on the mutation specific primers is guaranteed, (2) the modifiedblocker probe completely complementary to the wild type gene is introduced so that the wild type background interference is further reduced with keeping the efficiency of the mutant amplification, (3) based on the high specificity modified blocker probe, the ARMS primers can detect different base mutation forms at the same site at the same time so that multiple sites in the same gene can be detected by only a few systems,