FIG 1 RPCR-MC detection principle flow chart. From left to right, the sample was first extracted, and the sample and release agent were combined and mixed well. In the amplification stage, the initial sample is amplified by asymmetric amplification using upstream and downstream primers. Then, with the increase in temperature in the melting stage, the fluorescent groups and quenching groups of the probe become separated, and the binding ability of the probe to the template is reduced, resulting in a specific Tm value and forming a melting peak diagram. WT, wild-type; MT, mutant; W+M, heterozygosity.