Detection of PCB77 by Antibody-Coated Competitive Fluorescent Quantitative Immuno-PCR Using Molecular Beacon

Abstract

A rapid and quantitative technique is urgently needed in detecting toxicological and carcinogenic polychlorinated biphenyls (PCBs) in the environment. In this study, a high sensitive real time fluorescent quantitative immuno-PCR (FQ-IPCR) approach using molecular beacon (MB) was developed to detect PCB77, which was classified as a human carcinogen. MB-based FQ-IPCR was then performed on serial dilutions of known PCB77 concentrations equivalent to 10-fold dilutions of 10–10⁵ fg/mL. A correlation coefficient of 0.997 was identified, and a linear relationship between 10 fg/mL and 100 pg/mL, with y = 0.543x + 21.779, was obtained. Three soil samples were determined by MB-based FQ-IPCR to proof the validity of the method, and the results of which were further confirmed by conventional enzyme-linked immuno sorbent assay (ELISA). Based on sensitivity and reproduction, the MB-based real time FQ-IPCR technique will become a promising monitoring tool for environmental endocrine disruptors.

Keywords:

• Detection
• ELISA
• FQ-IPCR
• molecular beacon
• PCB77

This work was supported by the National Natural Science Foundation of China (20677008), the key project of fundamental research of Shanghai in China (09JC1407600), the Research Fund for the Doctoral Program of Higher Education (20060255044), and the Research Fund for the Doctoral Innovation Program of Donghua University (BC200832). We express our deep thanks.

Notes

RSD (n = 9): relative standard deviation.