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采用嵌入式电极的介电泳浓缩快速检测心肌标志物肌钙蛋白 I

Rapid electrical immunoassay of the cardiac biomarker troponin I through dielectrophoretic concentration using imbedded electrodes.

机构信息

School of Mechanical and Nuclear Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.

School of Mechanical and Nuclear Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea; Department of Biomedical Engineering, UNIST, Ulsan 44919, Republic of Korea; School of Materials Science and Engineering, UNIST, Ulsan 44919, Republic of Korea.

出版信息

Biosens Bioelectron. 2016 Aug 15;82:78-84. doi: 10.1016/j.bios.2016.03.056. Epub 2016 Mar 26.

Abstract

Rapidity and high sensitivity are critical factors for the diagnoses of heart attacks, and cardiac troponin I (cTnI) is at present a clinical standard for its diagnosis. Here we report a rapid, label-free, and highly sensitive single-walled carbon nanotube (SWCNT) electrical immunosensor, featuring two pairs of electrodes. Two concentration electrodes (gaps: 25 and 80µm) and two detection electrodes (source and drain; gap: 20µm; width: 50µm) were used for dielectrophoretic concentration of cTnI on the SWCNT channels and resistance measurements of the dielectrophoresis (DEP)-concentrated cTnI, respectively. The two concentration electrodes were imbedded between upper and lower dielectric layers, facing each other, underneath the -COOH-functionalized SWCNT channels deposited between the detection electrodes. Therefore, the gap between these imbedded concentration electrodes can be reduced to maximize the electric field intensity for DEP-mediated concentration of cTnI, thereby greatly reducing the detection time (1min) and enhancing the limit of detection (0.7-0.8pgmL(-)(1)). Relative resistance changes of the SWCNTs were measured as cTnI concentration in Tris-Borate-EDTA (TBE; 0.0025×) and human serum diluted 500-fold with 0.0025× TBE decreased from 100ngmL(-)(1) to 1pgmL(-1), and they were shown to be linear with the logarithm of cTnI concentration (R(2)=0.99 and 0.97, respectively). These immunosensors also showed high specificity over another cardiac biomarker, myoglobin, TBE medium (0.0025×), and 500-fold diluted human serum. The DEP-capture of cTnI depended on the frequency of the applied electric field, demonstrating the qualitative nature of the real part of the Clausius-Mossotti factor for cTnI.

摘要

快速和高灵敏度是诊断心脏病发作的关键因素,目前心肌肌钙蛋白 I (cTnI) 是其诊断的临床标准。在这里,我们报告了一种快速、无标记、高灵敏度的单壁碳纳米管 (SWCNT) 电化学生物传感器,具有两对电极。两个浓度电极(间隙:25 和 80µm)和两个检测电极(源和漏;间隙:20µm;宽度:50µm)用于在 SWCNT 通道上进行基于电介质电泳的 cTnI 浓缩以及对电介质电泳 (DEP)-浓缩 cTnI 的电阻测量。两个浓度电极被嵌入在上层和下层电介质层之间,彼此相对,位于置于检测电极之间的 -COOH 功能化 SWCNT 通道下方。因此,这些嵌入式浓度电极之间的间隙可以减小到最大限度地提高用于 DEP 介导的 cTnI 浓缩的电场强度,从而大大缩短检测时间(1 分钟)并提高检测限(0.7-0.8pgmL(-)(1))。当 cTnI 浓度在 Tris-Borate-EDTA (TBE;0.0025×) 和用 0.0025×TBE 稀释 500 倍的人血清中从 100ngmL(-)(1)降低到 1pgmL(-1)时,SWCNTs 的相对电阻变化得以测量,并且它们与 cTnI 浓度的对数呈线性关系(R(2)=0.99 和 0.97,分别)。这些免疫传感器在另一种心脏生物标志物肌红蛋白、TBE 介质(0.0025×)和 500 倍稀释的人血清中也表现出高特异性。cTnI 的 DEP 捕获取决于所施加电场的频率,证明了 Clausius-Mossotti 因子的实部对 cTnI 的定性性质。

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