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基于纳米粒子的免疫层析测试条与荧光探测器定量检测磷酸化乙酰胆碱酯酶:有机磷试剂的暴露生物标志物。

Nanoparticle-based immunochromatographic test strip with fluorescent detector for quantification of phosphorylated acetylcholinesterase: an exposure biomarker of organophosphorus agents.

机构信息

Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Science, Central China Normal University, Wuhan 430079, P R China.

出版信息

Analyst. 2013 Sep 21;138(18):5431-6. doi: 10.1039/c3an00621b. Epub 2013 Jul 25.

Abstract

A nanoparticle-based fluorescence immunochromatographic test strip (FITS) coupled with a hand-held detector for highly selective and sensitive detection of phosphorylated acetylcholinesterase (AChE), an exposure biomarker of organophosphate (OP) pesticides and nerve agents, is reported. In this approach, OP-AChE adducts were selectively captured by quantum dot-tagged anti-AChE antibodies (Qdot-anti-AChE) and zirconia nanoparticles (ZrO2 NPs). The sandwich-like immunoreactions were performed among the Qdot-anti-AChE, OP-AChE and ZrO2 NPs to form a Qdot-anti-AChE-OP-AChE-ZrO2 complex, which was detected by recording the fluorescence intensity of Qdots captured during the test line. Paraoxon was used as the model OP pesticide. Under optimal conditions, this portable FITS immunosensor demonstrates a highly linear absorption response over the range of 0.01 nM to 10 nM OP-AChE, with a detection limit of 4 pM, coupled with good reproducibility. Moreover, the FITS immunosensor has been validated with OP-AChE spiked human plasma samples. This is the first report on the development of ZrO2 NP-based FITS for the detection of the OP-AChE adduct. The FITS immunosensor provides a sensitive and low-cost sensing platform for on-site screening/evaluating OP pesticides and nerve agents poisoning.

摘要

一种基于纳米粒子的荧光免疫层析测试条(FITS)与手持式检测仪结合,用于高选择性和高灵敏度检测磷酸化乙酰胆碱酯酶(AChE),这是有机磷(OP)农药和神经毒剂的暴露生物标志物。在该方法中,OP-AChE 加合物被量子点标记的抗 AChE 抗体(Qdot-anti-AChE)和氧化锆纳米颗粒(ZrO2 NPs)选择性捕获。夹心式免疫反应在 Qdot-anti-AChE、OP-AChE 和 ZrO2 NPs 之间进行,形成 Qdot-anti-AChE-OP-AChE-ZrO2 复合物,通过记录测试线中捕获的 Qdot 的荧光强度来进行检测。对氧磷被用作模型 OP 农药。在最佳条件下,这种便携式 FITS 免疫传感器在 0.01 nM 至 10 nM OP-AChE 的范围内表现出高度线性的吸收响应,检测限为 4 pM,具有良好的重现性。此外,该 FITS 免疫传感器已通过添加 OP-AChE 的人血浆样本进行了验证。这是首次报道基于 ZrO2 NP 的 FITS 用于检测 OP-AChE 加合物。FITS 免疫传感器为现场筛选/评估 OP 农药和神经毒剂中毒提供了灵敏、低成本的传感平台。

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