A real-time fluorescent quantitative immuno-polymerase chain reaction （RT-IPCR） assay was developed for the detection of non-dioxin- like polychlorinated biphenyl （PCB） congener in soil samples. Based on the const...A real-time fluorescent quantitative immuno-polymerase chain reaction （RT-IPCR） assay was developed for the detection of non-dioxin- like polychlorinated biphenyl （PCB） congener in soil samples. Based on the construction of 3,4-dichlorinated biphenyl （IUPAC PCB 12） hapten and its immunogen, the specific polyclonal antibodies （pAbs） to PCB12 was obtained and used to develop a direct competitive RT-IPCR assay. Using the optimized assay, a standard curve for PCB 12 was prepared. The linear range for the determination of PCB 12 was from 10.0 to 1.0 x l06 fg/mL with a correlation coefficient of 0.98 and a detection limit of 1.53 fg/rnL. The RT-IPCR assays were tested for their cross-reactivity profiles using four selected congeners and four Aroclor products. The results for the soil samples correlated with the concentrations of PCBs obtained by gas chromatography/mass spectrometry. This highly specific, sensitive, and robust assay can be applied to on-site tests of PCBs and serve as a model for other pollutant immunoassays.展开更多
An α fetoprotein (AFP) antibody gene probe was constructed using chlorophyll molecule as a coupler between protein and dsDNA. The preliminary study on the detection of AFP using this novel probe was performed by immu...An α fetoprotein (AFP) antibody gene probe was constructed using chlorophyll molecule as a coupler between protein and dsDNA. The preliminary study on the detection of AFP using this novel probe was performed by immuno PCR, and the results indicated that the sensitivity of the gene probe by immuno PCR is 10 4-10 5 times higher compared with ELISA. The construction of immuno PCR gene probe in this way not only completely prevents the protein from contacting with organic solvent and maintains the native conformation of the proteins, but also anchors protein to dsDNA in a fixed orientation and makes PCR amplification more efficient. The gene probe thus constructed is stable for at least 6 months at room temperature. This new approach is exquisite, simple, less expensive, and suitable to a variety of applications.展开更多
基金supported by the National Natural Science Foundation of China (No. 21177082)the Science and Technology Commission of Shanghai Municipality in China (Key Project of Fundamental Research,No.09JC1407600)+1 种基金the Shanghai Jiao Tong University science and technology innovation special fund development projectsthe Postdoctoral Science Foundation of Shanghai (No. 11R21414600)
文摘A real-time fluorescent quantitative immuno-polymerase chain reaction （RT-IPCR） assay was developed for the detection of non-dioxin- like polychlorinated biphenyl （PCB） congener in soil samples. Based on the construction of 3,4-dichlorinated biphenyl （IUPAC PCB 12） hapten and its immunogen, the specific polyclonal antibodies （pAbs） to PCB12 was obtained and used to develop a direct competitive RT-IPCR assay. Using the optimized assay, a standard curve for PCB 12 was prepared. The linear range for the determination of PCB 12 was from 10.0 to 1.0 x l06 fg/mL with a correlation coefficient of 0.98 and a detection limit of 1.53 fg/rnL. The RT-IPCR assays were tested for their cross-reactivity profiles using four selected congeners and four Aroclor products. The results for the soil samples correlated with the concentrations of PCBs obtained by gas chromatography/mass spectrometry. This highly specific, sensitive, and robust assay can be applied to on-site tests of PCBs and serve as a model for other pollutant immunoassays.
文摘An α fetoprotein (AFP) antibody gene probe was constructed using chlorophyll molecule as a coupler between protein and dsDNA. The preliminary study on the detection of AFP using this novel probe was performed by immuno PCR, and the results indicated that the sensitivity of the gene probe by immuno PCR is 10 4-10 5 times higher compared with ELISA. The construction of immuno PCR gene probe in this way not only completely prevents the protein from contacting with organic solvent and maintains the native conformation of the proteins, but also anchors protein to dsDNA in a fixed orientation and makes PCR amplification more efficient. The gene probe thus constructed is stable for at least 6 months at room temperature. This new approach is exquisite, simple, less expensive, and suitable to a variety of applications.
