Premium
Accurate Zygote‐Specific Discrimination of Single‐Nucleotide Polymorphisms Using Microfluidic Electrochemical DNA Melting Curves
Author(s) -
Yang Allen H. J.,
Hsieh Kuangwen,
Patterson Adriana S.,
Ferguson B. Scott,
Eisenstein Michael,
Plaxco Kevin W.,
Soh H. Tom
Publication year - 2014
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201310059
Subject(s) - single nucleotide polymorphism , melting curve analysis , hybridization probe , high resolution melt , allele , genetics , genotype , snp , biology , dna , computational biology , microbiology and biotechnology , gene , polymerase chain reaction
Abstract We report the first electrochemical system for the detection of single‐nucleotide polymorphisms (SNPs) that can accurately discriminate homozygous and heterozygous genotypes using microfluidics technology. To achieve this, our system performs real‐time melting‐curve analysis of surface‐immobilized hybridization probes. As an example, we used our sensor to analyze two SNPs in the apolipoprotein E ( ApoE ) gene, where homozygous and heterozygous mutations greatly affect the risk of late‐onset Alzheimer’s disease. Using probes specific for each SNP, we simultaneously acquired melting curves for probe–target duplexes at two different loci and thereby accurately distinguish all six possible ApoE allele combinations. Since the design of our device and probes can be readily adapted for targeting other loci, we believe that our method offers a modular platform for the diagnosis of SNP‐based diseases and personalized medicine.