title: Developing a qPCR method to quantify AhR-PCP-DNA complex for detection of environmental trace-level PCP.
abstract: Pentachlorophenol (PCP), a widely-used aseptic or biocide, is known as an environmental toxicant involved in endocrine disruption even at a trace level. In order to reliably and efficiently quantify environmental trace-quantity PCP, this study developed a novel PCP detection method using the aryl hydrocarbon receptor (AhR) and fluorescence quantitative PCR (qPCR). DNA probe with AhR binding sites was synthesized by PCR before added into AhR-PCP complex. After AhR-PCP-DNA complex was digested with exonuclease, copy number of DNA probe was determined using fluorescence qPCR. To calculate PCP concentration in samples, a standard curve (PCP concentration versus Ct value) was constructed and the detection range was 10(-13) to 10(-9) M. PCP detection limit was 0.0089 ppt for the AhR-PCP-DNA complex assay and 8.8780 ppm for high performance liquid chromatography, demonstrating that the method developed in this study is more sensitive. These results suggest that AhR-PCP-DNA complex method may be successfully applicable in detection and quantification of environmental trace-level PCP.
authors.affiliation: College of Environmental Science and Engineering, Donghua University, Shanghai, 201620, People's Republic of China. zxx@dhu.edu.cn
authors.forename: Xiaoxiang
authors.initials: X
authors.lastname: Zhao
authors.forename: Xiaoqian
authors.initials: X
authors.lastname: Pang
authors.forename: Nuanapa
authors.initials: N
authors.lastname: Chaisuwan
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D015342:DNA Probes / Q000737:chemistry*; D004202:Disinfectants / Q000032:analysis* / Q000737:chemistry; D004784:Environmental Monitoring / Q000379:methods; D004785:Environmental Pollutants / Q000032:analysis* / Q000737:chemistry; D010416:Pentachlorophenol / Q000032:analysis* / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D018336:Receptors, Aryl Hydrocarbon / Q000737:chemistry*
chemical_list: D015342:DNA Probes; D004202:Disinfectants; D004785:Environmental Pollutants; D018336:Receptors, Aryl Hydrocarbon; D010416:Pentachlorophenol