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基于 Pt-Ir 纳米立方的去氢表雄酮侧向流免疫分析增强法

A Pt-Ir nanocube amplified lateral flow immunoassay for dehydroepiandrosterone.

机构信息

Department of Pharmaceutical Engineering, School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou 510006, People's Republic of China.

出版信息

Analyst. 2021 Apr 26;146(8):2726-2733. doi: 10.1039/d0an02293d.

Abstract

The traditional gold-nanoparticle-based lateral flow immunoassay (LFIA) cannot satisfy the requirements for the sensitive detection of dehydroepiandrosterone (DHEA) in human urine. To enhance the sensitivity of the LFIA, platinum-iridium nanocubes (Pt-Ir NCs) with high catalytic efficiency and stability were synthesized and labelled with polyclonal antibody (pAb) to form a pAb-Pt-Ir probe. For the detection of DHEA, a novel LFIA with Pt-Ir NCs as an optical label and an enhanced LFIA in which the peroxidase-like activity of the Pt-Ir NCs was triggered by the introduction of the chromogenic substrate 3-amino-9-ethyl-carbazole (AEC) were developed and compared with a LFIA with platinum nanocubes (PtNCs) as an optical label. The visual limit of detection was 0.5 ng mL-1 for Pt-Ir-LFIA and 0.05 ng mL-1 for AEC-enhanced Pt-Ir-LFIA, in comparison to 100 ng mL-1 for PtNCs-LFIA and 50 ng mL-1 for AEC-enhanced PtNCs-LFIA. The average recoveries from spiked urine samples ranged from 90.8% to 110.4%, with a coefficient of variation below 12.6%, suggesting the accuracy and reliability of our developed immunoassay. Achieving excellent sensitivity, specificity, and reproducibility, Pt-Ir-LFIA provided a promising platform for monitoring DHEA.

摘要

传统的基于金纳米颗粒的侧向流动免疫分析(LFIA)不能满足人体尿液中脱氢表雄酮(DHEA)灵敏检测的要求。为了提高 LFIA 的灵敏度,合成了具有高效催化稳定性的铂铱纳米立方体(Pt-Ir NCs),并用多克隆抗体(pAb)标记形成 pAb-Pt-Ir 探针。用于检测 DHEA,开发了一种新型 LFIA,其中 Pt-Ir NCs 用作光学标记,并且通过引入显色底物 3-氨基-9-乙基咔唑(AEC)触发 Pt-Ir NCs 的过氧化物酶样活性,与以铂纳米立方体(PtNCs)作为光学标记的 LFIA 进行了比较。Pt-Ir-LFIA 的视觉检测限为 0.5 ng mL-1,AEC 增强的 Pt-Ir-LFIA 的检测限为 0.05 ng mL-1,而 PtNCs-LFIA 的检测限为 100 ng mL-1,AEC 增强的 PtNCs-LFIA 的检测限为 50 ng mL-1。从尿液样本中添加的回收率平均在 90.8%至 110.4%之间,变异系数低于 12.6%,表明我们开发的免疫分析具有准确性和可靠性。Pt-Ir-LFIA 实现了出色的灵敏度、特异性和重现性,为监测 DHEA 提供了有前途的平台。

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