Tanaka Tsuyoshi, Hatakeyama Keiichi, Sawaguchi Masahiro, Iwadate Akihito, Mizutani Yasushi, Sasaki Kazuhiro, Tateishi Naofumi, Takeyama Haruko, Matsunaga Tadashi
Department of Biotechnology, Tokyo University of Agriculture & Technology, Koganei, Tokyo 184-8588, Japan.
Biotechnol Bioeng. 2006 Sep 5;95(1):22-8. doi: 10.1002/bit.20949.
A thin film transistor (TFT) photosensor fabricated by semiconductor integrated circuit (IC) technology was applied to DNA chip technology. The surface of the TFT photosensor was coated with TiO2 using a vapor deposition technique for the fabrication of optical filters. The immobilization of thiolated oligonucleotide probes onto a TiO2-coated TFT photosensor using gamma-aminopropyltriethoxysilane (APTES) and N-(gamma-maleimidobutyloxy) sulfosuccinimide ester (GMBS) was optimized. The coverage value of immobilized oligonucleotides reached a plateau at 33.7 pmol/cm2, which was similar to a previous analysis using radioisotope-labeled oligonucleotides. The lowest detection limits were 0.05 pmol/cm2 for quantum dot and 2.1 pmol/cm2 for Alexa Fluor 350. Furthermore, single nucleotide polymorphism (SNP) detection was examined using the oligonucleotide-arrayed TFT photosensor. A SNP present in the aldehyde dehydrogenase 2 (ALDH2) gene was used as a target. The SNPs in ALDH21 and ALDH22 target DNA were detected successfully using the TFT photosensor. DNA hybridization in the presence of both ALDH21 and ALDH22 target DNA was observed using both ALDH21 and ALDH22 detection oligonucleotides-arrayed TFT photosensor. Use of the TFT photosensor will allow the development of a disposable photodetecting device for DNA chip systems.
一种通过半导体集成电路(IC)技术制造的薄膜晶体管(TFT)光电传感器被应用于DNA芯片技术。采用气相沉积技术在TFT光电传感器表面涂覆TiO₂以制造光学滤光片。使用γ-氨丙基三乙氧基硅烷(APTES)和N-(γ-马来酰亚胺丁氧基)磺基琥珀酰亚胺酯(GMBS)将硫醇化寡核苷酸探针固定在涂覆TiO₂的TFT光电传感器上的方法得到了优化。固定化寡核苷酸的覆盖值在33.7 pmol/cm²时达到平台期,这与之前使用放射性同位素标记寡核苷酸的分析结果相似。量子点的最低检测限为0.05 pmol/cm²,Alexa Fluor 350的最低检测限为2.1 pmol/cm²。此外,使用寡核苷酸阵列TFT光电传感器检测了单核苷酸多态性(SNP)。醛脱氢酶2(ALDH2)基因中存在的一个SNP被用作靶点。使用TFT光电传感器成功检测到了ALDH21和ALDH22靶点DNA中的SNP。使用同时阵列有ALDH21和ALDH22检测寡核苷酸的TFT光电传感器观察到了在同时存在ALDH21和ALDH22靶点DNA时的DNA杂交。TFT光电传感器的使用将有助于开发用于DNA芯片系统的一次性光电检测设备。