National Institute of Advanced Industrial Science and Technology (AIST), 16-1 Onogawa, Tsukuba, Ibaraki, 305-8569, Japan.
National Institute of Advanced Industrial Science and Technology (AIST), 16-1 Onogawa, Tsukuba, Ibaraki, 305-8569, Japan.
Biosens Bioelectron. 2019 Jul 1;136:76-83. doi: 10.1016/j.bios.2019.04.047. Epub 2019 Apr 25.
We investigated the feasibility of simultaneous detection of multiple environmentally- and biomedically-relevant RNA biomarker target sequences on a single newly fabricated 384-ch sensor array chip aiming at practical application. The individual sensor is composed of a photolithographically-fabricated Au/Cr-based electrode modified with peptide nucleic acid (PNA) probes. The sensor array chips showed sequence-specific responses upon hybridization of the probes with target sequences complementary to the probes in contrast to mismatch versions. The target oligonucleotides have 15-22 mer sequences from messenger RNAs for estrogen-responsive genes and microRNAs for lung cancer biomarkers. The dependence on target concentrations of sensor responses was observed by using a single chip on which experiments for detection of several target concentrations proceeded simultaneously, with the detection limit of 7.33 × 10 M. As more realistic samples, oligonucleotide samples amplified by PCR from a synthesized template sequence were applied to the chip. They showed sequence-specific responses, revealing the potential for fabricated sensor array chips to be utilized to analyze PCR samples. Unlike complicated and expensive chips that require nanofabrication, our sensor array chips based on glass coated with gold thin films are simple and can be fabricated from inexpensive and readily available materials.
我们研究了在单个新制造的 384 通道传感器阵列芯片上同时检测多个环境和生物医学相关 RNA 生物标志物靶序列的可行性,旨在实现实际应用。每个传感器由经过光刻制造的基于金/铬的电极组成,该电极经过肽核酸 (PNA) 探针修饰。与错配版本相比,传感器阵列芯片在探针与与探针互补的靶序列杂交时表现出序列特异性反应。靶寡核苷酸具有来自雌激素反应基因的信使 RNA 和肺癌生物标志物的 microRNAs 的 15-22 个碱基序列。通过使用单个芯片同时进行几个目标浓度的检测实验,观察到传感器响应对目标浓度的依赖性,检测限为 7.33×10-7M。作为更现实的样本,从合成模板序列扩增的 PCR 寡核苷酸样品被应用于该芯片。它们表现出序列特异性反应,表明制造的传感器阵列芯片具有分析 PCR 样品的潜力。与需要纳米制造的复杂且昂贵的芯片不同,我们基于涂有金薄膜的玻璃的传感器阵列芯片简单,并且可以由廉价且易于获得的材料制造。