Department of Biotechnology, Motilal Nehru National Institute of Technology Allahabad, Prayagraj, 211004, U.P., India.
Biological Product Division, ICAR-IVRI Izatnagar, Bareilly, U.P., India.
Mikrochim Acta. 2024 Sep 30;191(10):636. doi: 10.1007/s00604-024-06666-w.
An aptamer-based electrochemical sensor for the sensitive detection of staphylococcal enterotoxin type A (SEA) is presented. The truncated aptamer AptSEA1.4 used in this work was screened using computational techniques, which reduced the cost of the SELEX screening process. The aptamer-SEA interactions were confirmed by employing circular dichroism (CD) and fluorescence spectroscopy. Afterwards, for developing an electrochemical aptasensor, a fabricated GNR/FTO aptasensor was prepared and characterized using scanning electron microscopy-energy-dispersive X-ray analysis (SEM-EDX), atomic force microscopy (AFM), cyclic voltammetry (CV), and square wave voltammetry (SWV). A detailed investigation of aptamer and SEA interaction in the presence of various experimental conditions was also conducted through SWV and electrochemical impedance spectroscopy (EIS). The aptamer exhibits a strong affinity for SEA, with a dissociation constant (Kd) of 19.93 nM. The aptasensor is sensitive, with a lower limit of detection of 12.44 pg mL. It has good stability, repeatability, and specificity and has displayed highly specific and sensitive detection SEA in spiked packaged mixed fruit juice and milk, with a recovery of 95-110%. The aptasensor has high promise for detecting SEA in other food items.
一种基于适体的电化学生物传感器,用于灵敏检测金黄色葡萄球菌肠毒素 A(SEA),被呈现出来。在这项工作中使用的截断适体 AptSEA1.4 是通过计算技术筛选出来的,这降低了 SELEX 筛选过程的成本。适体与 SEA 的相互作用通过使用圆二色性(CD)和荧光光谱学得到证实。之后,为了开发电化学适体传感器,制备了一个制造的 GNR/FTO 适体传感器,并通过扫描电子显微镜-能量色散 X 射线分析(SEM-EDX)、原子力显微镜(AFM)、循环伏安法(CV)和方波伏安法(SWV)进行了表征。还通过 SWV 和电化学阻抗谱(EIS)对各种实验条件下适体和 SEA 相互作用进行了详细研究。该适体对 SEA 具有很强的亲和力,解离常数(Kd)为 19.93 nM。该适体传感器具有很高的灵敏度,检测限低至 12.44 pg mL。它具有良好的稳定性、重复性和特异性,并在掺杂包装的混合果汁和牛奶中显示出对 SEA 的高度特异性和灵敏检测,回收率为 95-110%。该适体传感器在其他食品中检测 SEA 具有很高的应用前景。