Fluorescence Quantitative PCR For Detection Of Pentachlorophenol

Pentachlorophenol(PCP),one of the phenolic compounds,can interfere with the normal incretion function of animals,and it is also an important environmental hormone.Its chemical properties are relatively stable;it can accumulate in the sediments,and is difficult to be degraded.So the residue in the environment is relatively high.When it had been widely used as soil insecticide throughout the world, it caused seriously environmental problem.Up to now,many researchers have studied on the monitoring methods of PCP in and out the country,however,there are still no reports on analyzing and monitoring methods of detecting PCP using both fluorescence quantitative PCR technology and PCP receptor at home and abroad.So, investigation on this issue is concerned on in this paper.The main results obtained are as follows:1.Receptor which was extracted from goldfishes being domesticated in a certain concentration of PCP for a month,was used in the experiment.The agarose gel electrophoresis demonstrated that,its relative fluorescence intensity was stronger than the unexposed group.So the result showed that goldfishes could produce more related receptor under the existence of hormone,2.HAP was used to purify ligand-receptor complex in this study.The results indicated that this method could remove other proteins except PCP receptor,and the effect was notable.3.Conventional PCR and fluorescence quantitative PCR were performed using the ligand-receptor-DNA complex as template,which was used as qualitative analysis and quantitative analysis,respectively.The relationship between different concentrations of PCP and Ct values was obtained.The functional expression was: y=-0.448x+9.032.Analyzed results demonstrated that the linear relation was significant when the concentration of PCP was between 10^-13mol/L and 10^-9mol/L (R²=0.9975).The study also indicated that when the concentration of PCP was lower than 10^-13mol/L,the result was negative.Therefore the limit detection of the limit of detection was 10^-13mol/L.The repeatability of the results in fluorescence quantitative PCR was also proved good.4.The method of High Performance Liquid Chromatography(HPLC) was used in the experiment to compare with fluorescence quantitative PCR method.It was found that PCP could not be detected by this method when the concentration of was below10^-9mol/L,which further proved that the limit detection of fluorescence quantitative PCR was much lower.5.PCP in environmental samples were at last monitored by the fluorescence quantitative PCR method,and the results were acceptable. In conclusion,the method of fluorescence quantitative PCR detecting pentachlorophenol has been successfully developed in this experiment;this method has a lower detection limit,good reproducibility,and is easier to perform,so it can be severed as a novel method to detect trace pentachlorophenol in the environment rapidly and precisely.