Highly Sensitive and Multiplexed Detection of Low-Frequency Mutation in Fragmented ctDNA by a Dual-Role Mediator Blocker Amplification Strategy

's and colors. Multiplexed DMBA-MCA was developed to detect seven variants at 0.1-0.5% VAF in one tube. Our innovative method offers advantages including exceptional sensitivity, elimination of the requirement for specific fluorescence probes, shorter amplicons, and high multiplexing capacity, potentially revolutionizing clinical practice and precision medicine.