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Photoelectrochemical immunosensor for methylated RNA detection based on WS2 and poly(U) polymerase–triggered signal amplification
Microchimica Acta ( IF 5.3 ) Pub Date : 2020-10-09 , DOI: 10.1007/s00604-020-04572-5 Yue Wang 1 , Xi Fang 1 , Huanshun Yin 1 , Yunlei Zhou 1 , Yue Yang 2 , Shiyun Ai 1
Microchimica Acta ( IF 5.3 ) Pub Date : 2020-10-09 , DOI: 10.1007/s00604-020-04572-5 Yue Wang 1 , Xi Fang 1 , Huanshun Yin 1 , Yunlei Zhou 1 , Yue Yang 2 , Shiyun Ai 1
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A novel photoelectrochemical immunosensor has been constructed for the determination of methylated RNA. MoS2 nanosheets with large specific area were employed as photoactive material, gold nanoparticles were used as signal amplification unit and immobilization matrix of 4-mercaptophenylboronic acid, anti-m6A antibody was adopted as methylated RNA recognition reagent, and poly(U) polymerase–mediated RNA chain extension and Ru(NH3)63+ were used as assisted signal amplification unit. With the sensitization effect of Ru(NH3)63+, the photoactivity of WS2 nanosheets was improved greatly, which also improved the sensitivity. Using visible-light excitation and ascorbic acid as electron donor, the sensitive determination of methylated RNA was achieved by monitoring the photocurrent change with different concentrations of methylated RNA. This photoelectrochemical immunosensor has a wide linear relationship with methylated RNA concentration from 0.05 to 35 nM under optimal experimental conditions. The low detection limit of 14.5 pM was realized based on 3σ criterion. In addition to the good selectivity, this sensor also presents high reproducibility with a relative standard deviation of 1.4% for the photocurrent of seven electrodes. The applicability of the developed method was also investigated by detecting the level of methylated RNA in corn seedling leaves with and without sulfadiazine treatment. Graphical abstract A novel photoelectrochemical immunosensor was developed for methylated RNA detection using the photoactive material of MoS2 and poly(U) polymerase–mediated RNA chain extension. Graphical abstract A novel photoelectrochemical immunosensor was developed for methylated RNA detection using the photoactive material of MoS2 and poly(U) polymerase–mediated RNA chain extension.
中文翻译：
基于WS2和poly(U)聚合酶触发信号放大的甲基化RNA检测光电化学免疫传感器
已经构建了一种用于测定甲基化RNA的新型光电化学免疫传感器。采用比表面积大的MoS2纳米片作为光敏材料，金纳米粒子作为信号放大单元和4-巯基苯基硼酸的固定基质，抗m6A抗体作为甲基化RNA识别试剂，poly(U)聚合酶介导的RNA扩链和Ru(NH3)63+作为辅助信号放大单元。在Ru(NH3)63+的增感作用下，WS2纳米片的光活性大大提高，也提高了灵敏度。使用可见光激发和抗坏血酸作为电子供体，通过监测不同浓度甲基化 RNA 的光电流变化，实现了甲基化 RNA 的灵敏测定。这种光电化学免疫传感器在最佳实验条件下与甲基化 RNA 浓度具有广泛的线性关系，从 0.05 到 35 nM。基于 3σ 标准实现了 14.5 pM 的低检测限。除了良好的选择性外，该传感器还具有高重现性，七个电极的光电流的相对标准偏差为 1.4%。还通过检测有和没有磺胺嘧啶处理的玉米幼苗叶片中甲基化 RNA 的水平来研究所开发方法的适用性。图形摘要 使用 MoS2 的光活性材料和聚（U）聚合酶介导的 RNA 链延伸，开发了一种用于甲基化 RNA 检测的新型光电化学免疫传感器。
更新日期：2020-10-09
中文翻译：
基于WS2和poly(U)聚合酶触发信号放大的甲基化RNA检测光电化学免疫传感器
已经构建了一种用于测定甲基化RNA的新型光电化学免疫传感器。采用比表面积大的MoS2纳米片作为光敏材料，金纳米粒子作为信号放大单元和4-巯基苯基硼酸的固定基质，抗m6A抗体作为甲基化RNA识别试剂，poly(U)聚合酶介导的RNA扩链和Ru(NH3)63+作为辅助信号放大单元。在Ru(NH3)63+的增感作用下，WS2纳米片的光活性大大提高，也提高了灵敏度。使用可见光激发和抗坏血酸作为电子供体，通过监测不同浓度甲基化 RNA 的光电流变化，实现了甲基化 RNA 的灵敏测定。这种光电化学免疫传感器在最佳实验条件下与甲基化 RNA 浓度具有广泛的线性关系，从 0.05 到 35 nM。基于 3σ 标准实现了 14.5 pM 的低检测限。除了良好的选择性外，该传感器还具有高重现性，七个电极的光电流的相对标准偏差为 1.4%。还通过检测有和没有磺胺嘧啶处理的玉米幼苗叶片中甲基化 RNA 的水平来研究所开发方法的适用性。图形摘要 使用 MoS2 的光活性材料和聚（U）聚合酶介导的 RNA 链延伸，开发了一种用于甲基化 RNA 检测的新型光电化学免疫传感器。
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