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一种能够快速定量检测癌胚抗原的一体化双适体传感器。

All-in-one dual-aptasensor capable of rapidly quantifying carcinoembryonic antigen.

机构信息

Luminescent MD, LLC, Hagerstown, MD 21742, United States.

Luminescent MD, LLC, Hagerstown, MD 21742, United States.

出版信息

Biosens Bioelectron. 2017 Apr 15;90:46-52. doi: 10.1016/j.bios.2016.11.043. Epub 2016 Nov 16.

Abstract

Using a dual DNA aptamer (CEA aptamer linked to hemin aptamer), capable of rapidly capturing carcinoembryonic antigen (CEA) and hemin, an all-in-one dual-aptasensor with 1,1'-oxalyldiimidazole (ODI) chemiluminescence detection was developed for the early diagnosis of human cancer. CEA and hemin competitively bound with the dual DNA aptamer while the mixture in a detection cell was incubated for 30min at room temperature. When Amplex Red and HO were added in the detection cell after the incubation, the yield of resorufin formed from the reaction Amplex Red and HO depended on the concentration of HRP-mimicking G-quardruplex DNAzyme formed from the binding interaction between hemin and the dual DNA aptamer. Bright red light was observed with the addition of ODI and HO in the detection cell containing resorufin. Relative CL intensity of all-in-one dual-aptasensor, operated with the competitive reaction of CEA and hemin in the presence of the dual aptamer, was exponentially decreased with the increase of CEA concentration in human serum. The limit of detection (LOD=3σ) of the all-in-one dual-aptasensor which operated with excellent accuracy, precision, and reproducibility was as low as 0.58ng/ml. The good correlation between the easy to use all-in-one dual-aptasensor and conventional enzyme-linked immunosorbent assay (ELISA), operated with time consuming procedures (e.g., long incubations and multiple washings), indicates that the rapid all-in-one dual-aptasensor can be applied as a novel clinical tool for the early diagnosis of breast cancer.

摘要

利用能够快速捕获癌胚抗原(CEA)和血红素的双 DNA 适体(CEA 适体与血红素适体相连),开发了一种一体式双适体传感器,具有 1,1'-草酰二咪唑(ODI)化学发光检测功能,用于人类癌症的早期诊断。CEA 和血红素与双 DNA 适体竞争结合,而混合物在检测细胞中于室温下孵育 30 分钟。孵育后,在检测细胞中加入 Amplex Red 和 HO 时,形成的 Resorufin 的产量取决于从血红素与双 DNA 适体的结合相互作用形成的 HRP 模拟 G-四链体 DNA 酶与 Amplex Red 和 HO 的反应。在含有 Resorufin 的检测细胞中加入 ODI 和 HO 时,观察到明亮的红光。在存在双适体的情况下,CEA 和血红素的竞争性反应操作的一体式双适体传感器的相对 CL 强度随着人血清中 CEA 浓度的增加呈指数下降。操作精度高、精密度和重现性好的一体式双适体传感器的检测限(LOD=3σ)低至 0.58ng/ml。操作简便的一体式双适体传感器与传统酶联免疫吸附测定(ELISA)之间的良好相关性(例如,孵育时间长和多次洗涤)表明,快速的一体式双适体传感器可作为一种新型临床工具,用于早期诊断乳腺癌。

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