Construction of an electrochemical DNA chip for simultaneous genotyping of single nucleotide polymorphisms

• Published in: Analyst (London) 2005-01, Vol.130 (5), p.687-693
• Main Authors: TAKAHASHI, Masayoshi, OKADA, Jun, ITO, Keiko, HASHIMOTO, Michie, HASHIMOTO, Koji, YOSHIDA, Yuri, FURUICHI, Yasuhiro, OHTA, Yasuhiko, MISHIRO, Sunji, GEMMA, Nobuhiro
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Bisbenzimidazole; Chemistry; DNA - blood; Electrochemical methods; Electrochemistry - methods; Electrodes; Exact sciences and technology; Genotype; Humans; Oligonucleotide Array Sequence Analysis - methods; Oligonucleotide Probes; Polymorphism, Single Nucleotide - genetics
• ISSN: 0003-2654; 1364-5528
• DOI: 10.1039/b414030n

An electrochemical DNA chip was constructed for simultaneous genotyping of single nucleotide polymorphisms (SNPs) using genomic DNA extracted from blood samples. This chip consisted of electrodes located on a single piece of substrate and allele-specific oligonucleotide probes on the electrodes. As a first application, the 4 SNPs (MxA[-88], MxA[-123], MBL[X/Y], and MBL[A/B]), which have association with the efficacy of interferon therapy for HCV patient, were genotyped on the new DNA chip. Following hybridization of PCR products containing the 4 types of fragments, washing, bisbenzimide H33258 (Hoechst 33258) reaction and electrochemical analyses, 59 blood samples were genotyped by the chip method simultaneously. All procedures were completed within 2 h and the results were 100% concordant with those by the direct sequence method. The electrochemical DNA chip is expected to be a practical tool for SNPs genotyping.