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Reusable Silicon-Based Surface-Enhanced Raman Scattering Ratiometric Aptasensor with High Sensitivity, Specificity, and Reproducibility
Analytical Chemistry ( IF 6.7 ) Pub Date : 2017-09-21 00:00:00 , DOI: 10.1021/acs.analchem.7b01881 Huayi Shi 1 , Na Chen 1 , Yuanyuan Su 1 , Houyu Wang 1 , Yao He 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2017-09-21 00:00:00 , DOI: 10.1021/acs.analchem.7b01881 Huayi Shi 1 , Na Chen 1 , Yuanyuan Su 1 , Houyu Wang 1 , Yao He 1
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Rapid, sensitive, and accurate detection of adenosine triphosphate (ATP), the primary energy molecule, is critical for the elucidation of its unique roles in cell signaling and many cellular reactions. Up to date, a major challenge is still remaining for fabricating surface-enhanced Raman scattering (SERS) aptamer sensors (aptasensors) suitable for accurate and reliable quantification of ATP. Herein, we develop the ratiometric silicon SERS aptasensor for ATP detection, which is made of uniform silver nanoparticles (Ag NPs)-modified silicon wafer (Ag NPs@Si), followed by the functionalization with double-stranded DNA (dsDNA I). The dsDNA I is formed by the aptamer and its complementary DNA (cDNA), which contains two independent segments (e.g., 5′-Cy3-labeled DNA-C1, 3′-ROX-labeled DNA-C2). In the presence of ATP, ROX–DNA-C2 is dissociated from dsDNA I due to the formation of aptamer/ATP complex, leading to the attenuation of ROX signals, and meanwhile, Cy3 signals remain constant ascribed to the formation of dsDNA II caused by the supplementation of aptamer. As a result, ratiometric signals of the ratio of ROX intensity to Cy3 intensity (IROX/ICy3) can be achieved. Of particular significance, the developed ATP aptasensor features excellent reproducibility [e.g., the relative standard deviation (RSD) is less than ∼4%, comparable or superior to that of previously reported aptasensors], ultrahigh sensitivity [e.g., the detection of limit (LOD) reaches 9.12 pM, lower than that of other reported ATP SERS aptasensors], as well as good recyclability (e.g., ∼9.3% of RSD values of ratiometric signals within three cycles).
中文翻译：
具有高灵敏度，特异性和可重复性的可重复使用的基于硅的表面增强拉曼散射比例传感器
快速，灵敏和准确地检测主要能量分子三磷酸腺苷（ATP），对于阐明其在细胞信号传导和许多细胞反应中的独特作用至关重要。迄今为止，制造适用于准确和可靠地定量ATP的表面增强拉曼散射（SERS）适体传感器（aptasensors）仍然是一个主要挑战。本文中，我们开发了用于ATP检测的比例式硅SERS适体传感器，该传感器由均匀的银纳米颗粒（Ag NPs）修饰的硅片（Ag NPs @ Si）制成，然后用双链DNA（dsDNA I）进行功能化。dsDNA I由适体及其互补DNA（cDNA）形成，其包含两个独立的区段（例如5'-Cy3标记的DNA-C1、3'-ROX标记的DNA-C2）。在ATP存在的情况下，由于适体/ ATP复合物的形成，ROX–DNA-C2与dsDNA I分离，导致ROX信号衰减，同时，Cy3信号保持恒定，归因于适体的补充导致dsDNA II的形成。结果，ROX强度与Cy3强度之比的比例信号（可以实现I ROX / I Cy3）。特别重要的是，已开发的ATP适体传感器具有出色的重现性[例如，相对标准偏差（RSD）小于约4％，与以前报道的适体传感器相当或优于]，超高灵敏度[例如极限检测（LOD） ）达到9.12 pM，低于其他已报道的ATP SERS aptasensors []，并且具有良好的可回收性（例如，三个周期内比率信号的RSD值的9.3％左右）。
更新日期：2017-09-21
中文翻译：
具有高灵敏度，特异性和可重复性的可重复使用的基于硅的表面增强拉曼散射比例传感器
快速，灵敏和准确地检测主要能量分子三磷酸腺苷（ATP），对于阐明其在细胞信号传导和许多细胞反应中的独特作用至关重要。迄今为止，制造适用于准确和可靠地定量ATP的表面增强拉曼散射（SERS）适体传感器（aptasensors）仍然是一个主要挑战。本文中，我们开发了用于ATP检测的比例式硅SERS适体传感器，该传感器由均匀的银纳米颗粒（Ag NPs）修饰的硅片（Ag NPs @ Si）制成，然后用双链DNA（dsDNA I）进行功能化。dsDNA I由适体及其互补DNA（cDNA）形成，其包含两个独立的区段（例如5'-Cy3标记的DNA-C1、3'-ROX标记的DNA-C2）。在ATP存在的情况下，由于适体/ ATP复合物的形成，ROX–DNA-C2与dsDNA I分离，导致ROX信号衰减，同时，Cy3信号保持恒定，归因于适体的补充导致dsDNA II的形成。结果，ROX强度与Cy3强度之比的比例信号（可以实现I ROX / I Cy3）。特别重要的是，已开发的ATP适体传感器具有出色的重现性[例如，相对标准偏差（RSD）小于约4％，与以前报道的适体传感器相当或优于]，超高灵敏度[例如极限检测（LOD） ）达到9.12 pM，低于其他已报道的ATP SERS aptasensors []，并且具有良好的可回收性（例如，三个周期内比率信号的RSD值的9.3％左右）。
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