Institute of Medical Engineering, Department of Biophysics, School of Basic Medical Science, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, 710061, China.
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, 710061, China.
Talanta. 2022 Apr 1;240:123185. doi: 10.1016/j.talanta.2021.123185. Epub 2021 Dec 28.
Saxitoxin (STX), is one of the most dangerous and widespread paralytic shellfish toxins, causing a severe threat to the ecosystem and human health. So, it is important and highly essential to develop novel techniques for STX detection in a convenient, desirable, and low-cost manner. Herein, this study developed an electrolyte-insulator-semiconductor (EIS) sensor covered with a layer-by-layer prepared, positively-charged weak polyelectrolyte layer of poly (allylamine hydrochloride) (PAH) for the label-free detection of STX. The specific aptamer (Apt) sensitive to STX was electrostatically adsorbed onto the PAH layer. This leads to a preferentially flat orientation of the Apt within the Debye length, thus yielding a reduced charge-screening influence and a higher sensor signal. Each step of sensor surface modification, i.e. PAH adsorption, immobilization of Apt, and attachment of STX, was monitored by capacitance-voltage (C-V) and constant-capacitance (ConCap) measurements. Furthermore, atomic force microscopy (AFM) was employed to characterize the surface morphology and roughness of the PAH layer. Fluorescence microscopy was used to confirm the effective immobilization of Apt onto the PAH-modified EIS sensor. The results showed that the detection range of this aptasensor for STX detection was 0.5-100 nM and the detection limit was as low as 0.05 nM. Furthermore, this aptasensor showed good selectivity and 9 days' stability. The mussel tissue extraction test suggested that this aptasensor can be used to detect STX in real samples. This aptasensor provides a convenient approach for moderate, rapid, and label-free detection of marine biological toxins.
石房蛤毒素(STX)是最危险和分布最广泛的麻痹性贝类毒素之一,对生态系统和人类健康构成严重威胁。因此,开发一种方便、理想和低成本的新型 STX 检测技术非常重要和必要。在此,本研究开发了一种带有层层制备的带正电荷弱聚电解质层的电解质-绝缘体-半导体(EIS)传感器,用于无标记检测 STX。对 STX 敏感的特异性适体(Apt)通过静电吸附到 PAH 层上。这导致 Apt 在德拜长度内优先呈现平面取向,从而产生较小的电荷屏蔽影响和更高的传感器信号。传感器表面修饰的每一步,即 PAH 吸附、Apt 的固定化以及 STX 的附着,都通过电容-电压(C-V)和恒电容(ConCap)测量进行监测。此外,原子力显微镜(AFM)用于表征 PAH 层的表面形态和粗糙度。荧光显微镜用于确认 Apt 有效固定在 PAH 修饰的 EIS 传感器上。结果表明,该适体传感器检测 STX 的检测范围为 0.5-100 nM,检测限低至 0.05 nM。此外,该适体传感器表现出良好的选择性和 9 天的稳定性。贻贝组织提取试验表明,该适体传感器可用于检测实际样品中的 STX。该适体传感器为海洋生物毒素的中速、快速和无标记检测提供了一种便捷的方法。