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基于实时免疫-PCR 的灵敏免疫生物条码分析用于检测 3,4,3',4'-四氯联苯。

A sensitive immunosorbent bio-barcode assay based on real-time immuno-PCR for detecting 3,4,3',4'-tetrachlorobiphenyl.

机构信息

School of Environmental Science and Engineering, Shanghai Jiao Tong University, 800 Dong Chuan Road, Shanghai, 200240, China.

出版信息

Anal Bioanal Chem. 2014 Feb;406(6):1693-700. doi: 10.1007/s00216-013-7583-9. Epub 2014 Jan 24.

Abstract

A functionalized gold-nanoparticle bio-barcode assay, based on real-time immuno-PCR (IPCR), was designed for the determination of 3,4,3',4'-tetrachlorobiphenyl (PCB77). 15 nm gold nanoparticles were synthesized, and modified with thiol-capped DNA and goat anti-rabbit IgG. The nanoparticle probes were used to replace antibody-DNA conjugate in the IPCR, and were fixed on the PCR tube wall via the immune reaction. Real-time PCR was performed to quantify the DNA signal directly. Under optimized conditions, the new method was used to detect PCB77 with a linearity range from 5 pg L(-1) to 10 ng L(-1), and the limit of detection (LOD) was 1.72 pg L(-1). Real samples of Larimichthys polyactis, collected from the East China Sea, were analyzed. Recovery was from 82 % to 112 %, and the coefficient of variation (CV) was acceptable. The results were compared with GC-ECD, revealing that the method would be acceptable for providing rapid, semi-quantitative, and reliable test results for making environmental decisions.

摘要

一种基于实时免疫-PCR(IPCR)的功能化金纳米粒子生物条码分析方法被设计用于测定 3,4,3',4'-四氯联苯(PCB77)。合成了 15nm 的金纳米粒子,并将其用巯基封端的 DNA 和兔抗-山羊 IgG 修饰。纳米粒子探针用于替代 IPCR 中的抗体-DNA 缀合物,并通过免疫反应固定在 PCR 管壁上。直接进行实时 PCR 以定量 DNA 信号。在优化条件下,该新方法用于检测 PCB77 的线性范围为 5pgL(-1)至 10ngL(-1),检测限(LOD)为 1.72pgL(-1)。从东海采集的 Larimichthys polyactis 实际样品进行了分析。回收率在 82%至 112%之间,变异系数(CV)可接受。结果与 GC-ECD 进行了比较,表明该方法可用于快速、半定量和可靠地提供环境决策的测试结果。

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