Kadimisetty Karteek, Malla Spundana, Rusling James F
Department of Chemistry, University of Connecticut , Storrs, Connecticut 06269, United States.
Institute of Material Science , Storrs, Connecticut 06269, United States.
ACS Sens. 2017 May 26;2(5):670-678. doi: 10.1021/acssensors.7b00118. Epub 2017 May 2.
A novel, automated, low cost, three-dimensional (3-D) printed microfluidic array was developed to detect DNA damage from metabolites of chemicals in environmental samples. The electrochemiluminescent (ECL) detection platform incorporates layer-by-layer (LbL) assembled films of microsomal enzymes, DNA and an ECL-emitting ruthenium metallopolymer in ∼10 nm deep microwells. Liquid samples are introduced into the array, metabolized by the human enzymes, products react with DNA if possible, and DNA damage is detected by ECL with a camera. Measurements of relative DNA damage by the array assess the genotoxic potential of the samples. The array analyzes three samples simultaneously in 5 min. Measurement of cigarette and e-cigarette smoke extracts and polluted water samples was used to establish proof of concept. Potentially genotoxic reactions from e-cigarette vapor similar to smoke from conventional cigarettes were demonstrated. Untreated wastewater showed a high genotoxic potential compared to negligible values for treated wastewater from a pollution control treatment plant. Reactivity of chemicals known to produce high rates of metabolite-related DNA damage were measured, and array results for environmental samples were expressed in terms of equivalent responses from these standards to assess severity of possible DNA damage. Genotoxic assessment of wastewater samples during processing also highlighted future on-site monitoring applications.
开发了一种新型的、自动化的、低成本的三维(3-D)打印微流控阵列,用于检测环境样品中化学物质代谢产物造成的DNA损伤。该电化学发光(ECL)检测平台在约10纳米深的微孔中采用了微粒体酶、DNA和一种能发出ECL的钌金属聚合物的逐层(LbL)组装膜。将液体样品引入阵列,由人体酶进行代谢,若产物可能与DNA发生反应,则通过相机利用ECL检测DNA损伤。通过该阵列对相对DNA损伤的测量可评估样品的遗传毒性潜力。该阵列能在5分钟内同时分析三个样品。通过对香烟和电子烟烟雾提取物以及受污染水样的测量来验证概念。结果表明,电子烟蒸气可能产生与传统香烟烟雾类似的遗传毒性反应。与污染控制处理厂处理后的废水可忽略不计的值相比,未经处理的废水显示出较高的遗传毒性潜力。对已知会产生高比率与代谢产物相关的DNA损伤的化学物质的反应性进行了测量,并根据这些标准的等效反应来表示环境样品的阵列结果,以评估可能的DNA损伤的严重程度。对废水样品处理过程中的遗传毒性评估也凸显了未来的现场监测应用。