A sensitive immunosorbent bio-barcode assay based on real-time immuno-PCR for detecting 3,4,3',4'-tetrachlorobiphenyl.
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.
School of Environmental Science and Engineering, Shanghai Jiao Tong University, 800 Dong Chuan Road, Shanghai, 200240, China.
Guang-Xin
GX
Yang
Hui-Sheng
HS
Zhuang
Han-Yu
HY
Chen
Xian-Yin
XY
Ping
Dan
D
Bu
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000737:chemistry / Q000276:immunology; D055910:Antibodies, Immobilized / Q000737:chemistry / Q000276:immunology; D004247:DNA / Q000737:chemistry; D004784:Environmental Monitoring / Q000379:methods*; D006046:Gold / Q000737:chemistry*; D007118:Immunoassay / Q000379:methods; D007074:Immunoglobulin G / Q000276:immunology; D007164:Immunosorbents / Q000737:chemistry; D057230:Limit of Detection; D053758:Nanoparticles / Q000737:chemistry*; D010473:Perciformes / Q000378:metabolism; D011078:Polychlorinated Biphenyls / Q000032:analysis*; D011817:Rabbits; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013438:Sulfhydryl Compounds / Q000737:chemistry; D014874:Water Pollutants, Chemical / Q000032:analysis*
D000888:Antibodies, Anti-Idiotypic; D055910:Antibodies, Immobilized; D007074:Immunoglobulin G; D007164:Immunosorbents; D013438:Sulfhydryl Compounds; D014874:Water Pollutants, Chemical; D006046:Gold; D004247:DNA; D011078:Polychlorinated Biphenyls; C028451:3,4,3',4'-tetrachlorobiphenyl