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Nucleic Acids Research 2004 32(18):e141; doi:10.1093/nar/gnh141
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Published online 21 October 2004

Nucleic Acids Research, Vol. 32 No. 18 © Oxford University Press 2004; all rights reserved

A novel method of identifying genetic mutations using an electrochemical DNA array

Junko Wakai, Atsuko Takagi1, Masato Nakayama, Takahito Miya, Takatoshi Miyahara, Tsuyoshi Iwanaga2, Shigeori Takenaka3, Yasuyuki Ikeda2 and Masahiko Amano*

TUM Gene, Inc., 3-1 Kazusa-Koito Kimitsu, Chiba 292-1149 Japan, 1 Department of Pharmacology and 2 Departments of Etiology and Pathophysiology, National Cardiovascular Center Research Institute, Fujishirodai, Suita, Osaka 565-8565 Japan and 3 Department of Applied Chemistry, Faculty of Engineering, Kyushu University, Fukuoka 812-8581, Japan

* To whom correspondence should be addressed. Tel: +81 439 70 1321; Fax: +81 439 32 5689; Email: amano{at}tum-gene.co.jp

Received July 3, 2004; Revised September 15, 2004; Accepted September 28, 2004

We describe the development of a new type of DNA array chip that utilizes electrochemical reactions and a novel method of simultaneously identifying multiple genetic mutations on an array chip. The electrochemical array (ECA) uses a threading intercalator specific to double-stranded nucleotides, ferrocenylnaphthalene diimide (FND), as the indicator. ECA does not require target labeling, and the equipment is simple, durable and less expensive. The simultaneous multiple mutation detection (SMMD) system using an ECA chip and FND utilizes an enzyme to simultaneously distinguish several genetic mutations such as single nucleotide polymorphism (SNP), insertion, deletion, translocation and short tandem repeat. We examined this SMMD system using an ECA chip, by detecting seven different mutations on the lipoprotein lipase (LPL) gene for 50 patients in a blind test. It turned out that all the results obtained were concordant with the sequencing results, demonstrating that this system is a powerful tool for clinical applications.


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