PARProceedings of Anticancer Research2208-35452208-3553Bio-Byword Scientific Publishing Pty. Ltd.10.26689/par.v4i6.1674Articlehttps://artdesignp.com/journal/PAR/4/6/10.26689/par.v4i6.1674ZhangJinku,SunJirui,QiaoHaizhi,HanLu,LiuYunjia2020462020-12-28Objective: To explore a method to solve the issue of interference in fluorescence quantitative PCR non-specific amplification for gene detection. Method: A three-step method was used for amplification, and the quantitative fluorescence signal collection process was set in the extension stage. Results: Three-step amplification has the advantages of wide application range; improved accuracy; and reduced primer design requirements. Conclusion: The interference of non-specific amplification signals was effectively avoided, the melting curve plotting process was omitted, the reaction time was shortened, and the detection accuracy was improved.