Wakai Junko, Takagi Atsuko, Nakayama Masato, Miya Takahito, Miyahara Takatoshi, Iwanaga Tsuyoshi, Takenaka Shigeori, Ikeda Yasuyuki, Amano Masahiko, Urata Tomohiro
TUM Gene, Inc., 3-1 Kazusa-Koito Kimitsu, Chiba 292-1149 Japan.
Nucleic Acids Res. 2004 Oct 21;32(18):e141. doi: 10.1093/nar/gnh141.
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.
我们描述了一种新型DNA阵列芯片的开发,该芯片利用电化学反应以及一种在阵列芯片上同时识别多种基因突变的新方法。电化学阵列(ECA)使用一种对双链核苷酸特异的穿线嵌入剂二茂铁基萘二亚胺(FND)作为指示剂。ECA不需要对靶标进行标记,且设备简单、耐用且成本较低。使用ECA芯片和FND的同时多重突变检测(SMMD)系统利用一种酶同时区分几种基因突变,如单核苷酸多态性(SNP)、插入、缺失、易位和短串联重复。我们通过在一项盲测中对50名患者脂蛋白脂肪酶(LPL)基因上的七种不同突变进行检测,使用ECA芯片对该SMMD系统进行了检验。结果表明,所有获得的结果与测序结果一致,证明该系统是临床应用的有力工具。