4.7 Article

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

Journal

ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 406, Issue 6, Pages 1693-1700

Publisher

SPRINGER HEIDELBERG
DOI: 10.1007/s00216-013-7583-9

Keywords

Gold nanoparticle; PCB77; Real-time immuno-PCR; Barcode DNA

Funding

  1. National Natural Science Foundation of China [21177082]
  2. Science and Technology Commission of Shanghai Municipality in China [09JC1407600]
  3. Shanghai Jiaotong University
  4. Shanghai Jiao Tong University

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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.

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