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Rapid Screening for EGFR Mutations Based on Selective Fluorescence Quenching of Gold Nanoparticles

H.X. Li, S. Hu and L. Rothberg
University of Rochester, US

Keywords:
rapid screening, egfr mutation, fluorescence quenching, gold nanoparticle

Abstract:
Here we report a simple, rapid and cost effective screening method for egfr mutations in PCR samples. This method is derived from a series of new observations: single strand DNA adsorbs to negatively charged gold nanoparticle (Au-np), and the adsorption is sequence length and temperature dependent while double strand DNA does not or much slowly adsorbs to Au-np. We use rhodamine red labeled short DNA sequence as probes. If the short probe hybridizes with target, it will not adsorb to Au-np, the probe fluoresces. Otherwise the probe absorbs to Au-np and the fluorescence is quenched because the probe does not hybridize with target and stays in single stranded. By choosing proper annealing temperature, we can differentiate perfect match or single base mismatch between the probe and target through fluorescence quenching. This assay only uses commercial available cheap reagents and takes less than 10 minutes. We detected 20 black samples and get 100% accuracy. We also get very striking mutation signal from poorly amplified PCR samples.

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