Agar Meltem, Laabei Maisem, Leese Hannah S, Estrela Pedro
Department of Electronic and Electrical Engineering, University of Bath, Bath, BA2 7AY, United Kingdom; Centre for Bioengineering & Biomedical Technologies (CBio), University of Bath, Bath, BA2 7AY, United Kingdom.
Department of Life Sciences, University of Bath, Bath, BA2 7AY, United Kingdom; School of Cellular and Molecular Medicine, University of Bristol, Bristol, BS8 1TD, United Kingdom.
Biosens Bioelectron. 2025 Mar 15;272:117136. doi: 10.1016/j.bios.2025.117136. Epub 2025 Jan 4.
Bacteria pose a significant threat to human health as they can cause diseases and outbreaks; therefore rapid, easy, and specific detection of bacteria in a short time is crucial. Various methods such as polymerase chain reaction and enzyme-linked immunosorbent assay have been developed for bacteria detection. However, most of these methods require sample preparation, trained personnel, and 2-4 days for identification. In this study, an electrochemical sensor has been developed in which a molecularly imprinted polymer (MIP) and aptamer were used together as a bioreceptor for the multiplexed detection of Staphylococcus aureus and Escherichia coli. Non-Faradaic electrochemical impedance spectroscopy (EIS) was employed to assess bacterial detection. Sensor performance was assessed in buffer solution, deionized water and spiked tap water. Aptamer-molecularly imprinted polymer (Apta-MIP) based electrochemical sensors demonstrate high sensitivity and selectivity for the detection of S. aureus and E. coli, with limits of detection of 4 CFU/mL and 2 CFU/mL, respectively. Additionally, these sensors exhibited a broad dynamic range from 1 CFU/mL to 10 CFU/mL. The Apta-MIPs performance surpasses those obtained for Aptasensors alone and MIPs alone, demonstrating the high efficiency of the double recognition effect that originates from the affinity between aptamer and bacteria and target-specific cavities on the polymer. This is the first study in which aptamers and imprinted polymers were used as a hybrid bioreceptors for multiplexed detection of bacteria. The Apta-MIP sensors produced in this study can be used as a point-of-care diagnostic tool for bacteria-related diseases and test of water quality.
细菌对人类健康构成重大威胁，因为它们可引发疾病和疫情；因此，在短时间内快速、简便且特异性地检测细菌至关重要。已开发出多种方法用于细菌检测，如聚合酶链反应和酶联免疫吸附测定。然而，这些方法大多需要样品制备、专业人员，且鉴定需要2至4天。在本研究中，开发了一种电化学传感器，其中分子印迹聚合物（MIP）和适配体一起用作生物受体，用于金黄色葡萄球菌和大肠杆菌的多重检测。采用非法拉第电化学阻抗谱（EIS）评估细菌检测情况。在缓冲溶液、去离子水和加标自来水体系中评估传感器性能。基于适配体 - 分子印迹聚合物（Apta - MIP）的电化学传感器对金黄色葡萄球菌和大肠杆菌的检测具有高灵敏度和选择性，检测限分别为4 CFU/mL和2 CFU/mL。此外，这些传感器在1 CFU/mL至10 CFU/mL范围内表现出较宽的动态范围。Apta - MIP的性能超过单独的适配体传感器和单独的MIP，证明了源自适配体与细菌之间的亲和力以及聚合物上的靶标特异性空腔的双重识别效应的高效性。这是首次将适配体和印迹聚合物用作混合生物受体进行细菌多重检测的研究。本研究中制备的Apta - MIP传感器可作为与细菌相关疾病和水质检测的即时诊断工具。
