关键词: 链置换反应, 金属增强荧光, 碱基突变筛查

Abstract: Objective To develop a highly sensitive and selective method for mutation detection utilizing metal enhanced fluorescence (MEF). Methods Strand-displacement was initiated by introducing the target ssDNA and resulting in the unfolding of the capture probe. A signal probe coupled with Carboxy fluorescein(FAM) was further hybridizing with the unfolding capture probe and generating fluorescent signal. Results Comparing to normal method, the MEF design could discriminate complementary target from targets with mismatch, and holds noticeable specificity and highly sensitivity (~13 fold signal amplification with 100nmol/L target) with the detection limit of 0.1nmol/L in silver-coated 96-well plate (control group is 5nmol/L). Conclusion The MEF assay provided an alternative screening method for mutation detection.

Key words: strand-displacement reaction, metal-enhanced fluorescence, mismatch screening