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
The gastric cancer associated antigen MG_7-Ag was detected by means of a newly established method,termed Immuno-PCR A McAb-recombinant DNA chimeric molecule was made which possesses bispecific binding affinity for ant...The gastric cancer associated antigen MG_7-Ag was detected by means of a newly established method,termed Immuno-PCR A McAb-recombinant DNA chimeric molecule was made which possesses bispecific binding affinity for antigen that had been immobilized on microtiter wells and the segment of the attached DNA was amplified by PCR. The antigen of gastric cancer cell line KATO III was monitored by this method. Analysis of PCR products by agarose gel electrophoresis after staining with ethidium bromide allowed as few as 20 cells to be detected readily and reproducibly. Immuno-PCR showed a 104 enhancement in detection sensitivity compared with ELISA assay. When the same numbers of cells ( 2×10e/ml ) were immobilized and then the serial diluted chimeric molecule was added. 3.8×10-14 moles and 3.0 × 10-11 moles were needed to give positive results with the Immuno-PCR and ELISA assay. respectively.Therefore, Immuno-PCR could give an enomous amplification capability with good specificity, and has a sensitivity much higher than any existing techniques for antigen detection.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The gastric cancer associated antigen MG_7-Ag was detected by means of a newly established method,termed Immuno-PCR A McAb-recombinant DNA chimeric molecule was made which possesses bispecific binding affinity for antigen that had been immobilized on microtiter wells and the segment of the attached DNA was amplified by PCR. The antigen of gastric cancer cell line KATO III was monitored by this method. Analysis of PCR products by agarose gel electrophoresis after staining with ethidium bromide allowed as few as 20 cells to be detected readily and reproducibly. Immuno-PCR showed a 104 enhancement in detection sensitivity compared with ELISA assay. When the same numbers of cells ( 2×10e/ml ) were immobilized and then the serial diluted chimeric molecule was added. 3.8×10-14 moles and 3.0 × 10-11 moles were needed to give positive results with the Immuno-PCR and ELISA assay. respectively.Therefore, Immuno-PCR could give an enomous amplification capability with good specificity, and has a sensitivity much higher than any existing techniques for antigen detection.
