A reliable and sensitive competitive real-time fluorescent quantitative immuno-PCR （RTFQ-IPCR） assay using a molecular beacon was developed for the determination of trace fluoranthene （FL） in the environment.Under...A reliable and sensitive competitive real-time fluorescent quantitative immuno-PCR （RTFQ-IPCR） assay using a molecular beacon was developed for the determination of trace fluoranthene （FL） in the environment.Under optimized assay conditions,FL can be determined in the concentration range from 1 fg/mL to 100 ng/mL,with y=0.194x ＋ 7.859,and a correlation coefficient of 0.967 was identified,with a detection limit of 0.6 fg/mL.Environmental water samples were successfully analyzed,recovery was between 90% and 116%,with intra-day relative standard deviation （RSD） of 6.7%-12.8% and inter-day RSD of 8.4%-15.2%.The results obtained from RTFQ-IPCR were confirmed by ELISA,showing good accuracy and suitability to analyze FL in field samples.As a highly sensitive method,the molecular beacon-based RTFQ-IPCR is acceptable and promising for providing reliable test results to make environmental decisions.展开更多
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
基金support by the Scienceand Technology Commission of Shanghai Municipality in China (Key Project of Fundamental Research) (No.09JC1407600)the Science and Technology Commission of Shanghai Municipality in China (Key Project of theScience and Technology Research) (No. 09231202805)the Shanghai Leading Academic Discipline Project(No. B604)
文摘A reliable and sensitive competitive real-time fluorescent quantitative immuno-PCR （RTFQ-IPCR） assay using a molecular beacon was developed for the determination of trace fluoranthene （FL） in the environment.Under optimized assay conditions,FL can be determined in the concentration range from 1 fg/mL to 100 ng/mL,with y=0.194x ＋ 7.859,and a correlation coefficient of 0.967 was identified,with a detection limit of 0.6 fg/mL.Environmental water samples were successfully analyzed,recovery was between 90% and 116%,with intra-day relative standard deviation （RSD） of 6.7%-12.8% and inter-day RSD of 8.4%-15.2%.The results obtained from RTFQ-IPCR were confirmed by ELISA,showing good accuracy and suitability to analyze FL in field samples.As a highly sensitive method,the molecular beacon-based RTFQ-IPCR is acceptable and promising for providing reliable test results to make environmental decisions.
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.
