Developing a qPCR method to quantify AhR–PCP–DNA complex for detection of environmental trace-level PCP

摘要

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.