Institute for Environmental Monitoring and Evaluation, School of Environmental Science and Engineering, Shanghai Jiao Tong University, Shanghai 200240, China.
Acta Biochim Biophys Sin (Shanghai). 2018 Apr 1;50(4):381-390. doi: 10.1093/abbs/gmy020.
Benzo[a]pyrene (BaP), a type of polycyclic aromatic hydrocarbon, is widely distributed in the environment. In this study, BaP immunogen and coating antigen were respectively prepared by different methods, and the specific antibody targeting the BaP analyte was obtained. Moreover, gold nanoparticles modified with the specific polyclonal antibody and thiol-capped DNA were prepared as biological probes. Based on the work above, an ultrasensitive biological probe enhanced real-time immuno-polymerase chain reaction (BP-rt-iPCR) assay was developed to detect BaP in several environmental samples. Under optimal conditions, the proposed method was used to detect BaP with a linearity ranging from 5 pg/l to 50 ng/l. The limit of detection was 1.63 pg/l. The recovery rates of the spiked samples ranged from 92.12% to 109.76% and the coefficients of variation were 7.53%-11.06%. This immunoassay was successfully used to detect BaP in environmental samples, and the BaP detection results were consistent with those obtained using high performance liquid chromatography, indicating that the BP-rt-iPCR method is accurate and reliable, and has great potential to detect trace amounts of BaP.
苯并[a]芘（BaP）是一种多环芳烃，广泛分布于环境中。本研究分别采用不同方法制备 BaP 免疫原和包被抗原，获得了针对 BaP 分析物的特异性抗体。此外，还制备了金纳米粒子修饰的特异性多克隆抗体和巯基封端的 DNA 作为生物探针。基于上述工作，开发了一种超灵敏的生物探针增强实时免疫聚合酶链反应（BP-rt-iPCR）测定法，用于检测几种环境样品中的 BaP。在最佳条件下，该方法用于检测 BaP 的线性范围为 5 pg/l 至 50 ng/l，检测限为 1.63 pg/l。加标样品的回收率范围为 92.12%至 109.76%，变异系数为 7.53%-11.06%。该免疫测定法成功地用于检测环境样品中的 BaP，BaP 的检测结果与高效液相色谱法一致，表明 BP-rt-iPCR 方法准确可靠，具有检测痕量 BaP 的巨大潜力。
