“…Many efforts have been devoted to multiplex SNV discrimination and detection, for example, DNA sequencing, , polymerase chain reaction (PCR), − the microarray, − enzyme-assisted methods, − and hybridization-based methods, − where the discrimination readout can be optical, − electrical, mass spectrometric, or electrochemical. ,,,− However, limited approaches can realize multiplex SNV detection with both ultrahigh specificity and sensitivity in a simple manner. To achieve the high discrimination factor and low-abundance detection capability, most available methods require an elaborate and sophisticated probe or primer design, time-consuming procedures, and the tedious amplification step in the workflow. ,,,− In the efforts toward enzyme-free, highly specific SNV discrimination, the emergence of competitive or masking systems sheds lights on the development of highly specific hybridization-based SNV discrimination technologies. ,,,,,, Among the masking or sequester hairpin is an extremely simple masking reagent that enables significantly enhanced SNV discrimination without complicating probe designs for detection, because the hairpin structure will not interfere with the other probes in the test system; in combination with the molecular beacon for signal recognition and signal readout, the question is high discrimination factors (>200) were not consistently achieved for the majority of all tested SNVs. Also, the requirement for dual labeling of the different fluorophore and quencher increases the reagent cost .…”