384-Channel electrochemical sensor array chips based on hybridization-triggered switching for simultaneous oligonucleotide detection
384-Channel electrochemical sensor array chips based on hybridization-triggered switching for simultaneous oligonucleotide detection
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DOI：
10.1016/j.bios.2019.04.047
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发表时间：
2019-07-01
影响因子：
12.6
通讯作者：
Nakazato, Tetsuya
中科院分区：
文献类型：
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作者：
Aoki, Hiroshi;Torimura, Masaki;Nakazato, Tetsuya
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，旨在实际应用。单个传感器由光刻制造的 Au/Cr 基电极组成，并用肽核酸 (PNA) 探针进行修饰。与错配版本相比，传感器阵列芯片在探针与与探针互补的目标序列杂交时显示出序列特异性响应。目标寡核苷酸具有 15-22 mer 序列，来自雌激素响应基因的信使 RNA 和肺癌生物标志物的 microRNA。通过使用单芯片观察传感器响应对目标浓度的依赖性，在该芯片上同时进行多个目标浓度的检测实验，检测限为7.33 x 10(-8) M。作为更真实的样品，将通过PCR从合成的模板序列扩增的寡核苷酸样品应用于芯片。他们表现出序列特异性反应，揭示了制造的传感器阵列芯片用于分析 PCR 样本的潜力。与需要纳米加工的复杂而昂贵的芯片不同，我们的基于涂有金薄膜的玻璃的传感器阵列芯片很简单，并且可以用廉价且容易获得的材料来制造。
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 x 10(-8) 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.