title: Single-nucleotide polymorphism genotyping using a novel multiplexed electrochemical biosensor with nonfouling surface.
abstract: A novel electrochemical DNA biosensor for single-nucleotide polymorphism (SNP) analysis was developed. In this work, an oligonucleotide-incorporated nonfouling surface (ONS) was constructed to resist nonspecific absorption. The biosensor was developed using a 16-electrode array for high-throughput SNP analysis. The proposed strategy was primarily based on specific oligonucleotide ligation. Fully matched target DNA templated the ligation between a capture probe assembled on gold electrodes and a tandem signal probe with a biotin moiety that could capture avidin-horseradish peroxidase and sequentially generate a catalysed amperometric signal. A pre-core mutation in the hepatitis B virus (HBV) genome at G1896A and two adjacent polymorphisms in the human CYP2C19 genome at C680T and G681A were analysed. Polymerase chain reaction (PCR) products were used as real-life samples and analysed. Our results showed that 10% of a single-mismatched mutant gene was clearly distinguished with a current signal 16 times higher than that of the blank sample, demonstrating the selectivity and practicability of the multiplexed electrochemical DNA biosensor.
authors.affiliation: Division of Chemistry and Ionizing Radiation Measurement Technology, Shanghai Institute of Measurement and Testing Technology, Shanghai 201203, PR China.
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
authors.forename: Ruojun
authors.initials: R
authors.lastname: Lao
authors.forename: Li
authors.initials: L
authors.lastname: Xu
authors.forename: Qin
authors.initials: Q
authors.lastname: Xu
authors.forename: Lanying
authors.initials: L
authors.lastname: Li
authors.forename: Min
authors.initials: M
authors.lastname: Zhang
authors.forename: Shiping
authors.initials: S
authors.lastname: Song
authors.forename: Chunhai
authors.initials: C
authors.lastname: Fan
mesh_terms: D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics*; D015374:Biosensing Techniques / Q000379:methods*; D065731:Cytochrome P-450 CYP2C19; D055664:Electrochemical Techniques / Q000379:methods; D060005:Genotyping Techniques; D006515:Hepatitis B virus / Q000235:genetics*; D057166:High-Throughput Screening Assays; D006801:Humans; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D013499:Surface Properties
chemical_list: D001189:Aryl Hydrocarbon Hydroxylases; C045793:CYP2C19 protein, human; D065731:Cytochrome P-450 CYP2C19