Fan Ziyan, Li Zhonghao, Liu Shanshan, Yang Fei, Bian Zhaoyang, Wang Ying, Tang Gangling, Zhao Qinxiao, Deng Huimin, Liu Shili
China National Tobacco Quality Supervision and Test Center No. 2 Fengyang Street, High and New Technology Industries Development Zone Zhengzhou 450001 China
School of Basic Medical Science, Shandong University Jinan Shandong 250012 China.
RSC Adv. 2018 Aug 21;8(52):29562-29569. doi: 10.1039/c8ra04915g. eCollection 2018 Aug 20.
Benzo[]pyrene (BaP) is considered as one of the most carcinogenic pollutants in cigarette smoke. The development of simple and sensitive BaP screening methods can help assess the risk of cigarette exposure to the human body rapidly. In this report, a rapid fluorescence immunoassay (RFIA) method for the detection of BaP is proposed, the core of which is the synthesis of bifunctional covalent antibody-DNA conjugates for target recognition and signal amplification. Based on the optimization of the SYBR Green I and PAH-BSA concentrations, as well as DNA-antibody immune complex's dilution in the RFIA system, a serial dilution of BaP was tested with this method. The results showed that the linear working range of the RFIA for BaP is 0.46 to 333 ng mL, which is much wider than traditional ELISA. The detection limit was 0.32 ng mL, which was more sensitive than other methods such as the redox-labeled electrochemical immunoassay method and the competitive piezoelectric biosensor. Then the cross-reactions (CR) of other PAHs in cigarette smoke were evaluated using this RFIA and found that the cross-reactions of naphthalene, anthracene, and pyrene were very low (<1%). The cross-reaction in this RFIA system can be reduced by improving the specificity of the antibody. To the best of our knowledge, this is the first time that the BaP in mainstream cigarette smoke was tested; the RFIA demonstrates fast and simple experimental manipulations and better working curves and sensitivity.
苯并[a]芘（BaP）被认为是香烟烟雾中最具致癌性的污染物之一。开发简单灵敏的BaP筛查方法有助于快速评估香烟暴露对人体的风险。在本报告中，提出了一种用于检测BaP的快速荧光免疫分析（RFIA）方法，其核心是合成用于目标识别和信号放大的双功能共价抗体 - DNA缀合物。基于对SYBR Green I和PAH - BSA浓度以及RFIA系统中DNA - 抗体免疫复合物稀释度的优化，用该方法对BaP的系列稀释液进行了测试。结果表明，BaP的RFIA线性工作范围为0.46至333 ng/mL，比传统酶联免疫吸附测定（ELISA）宽得多。检测限为0.32 ng/mL，比氧化还原标记电化学免疫分析方法和竞争性压电生物传感器等其他方法更灵敏。然后用该RFIA评估了香烟烟雾中其他多环芳烃（PAHs）的交叉反应（CR），发现萘、蒽和芘的交叉反应非常低（<1%）。该RFIA系统中的交叉反应可通过提高抗体的特异性来降低。据我们所知，这是首次对主流香烟烟雾中的BaP进行测试；RFIA显示出快速简单的实验操作以及更好的工作曲线和灵敏度。
