基于适配体的阻抗传感器用于细菌分型
Aptamer-based Impedimetric Sensor for Bacterial Typing
摘要：
介绍了一种基于适体的阻抗式细菌分型传感器（AIST-B）的研制。通过Cell-SELEX技术筛选出对沙门氏菌具有高度特异性的DNA适体。进行了12轮选择;每轮包括针对S的阳性选择步骤。针对相关病原体的混合物，包括鼠伤寒沙门氏菌、大肠杆菌、金黄色葡萄球菌、铜绿假单胞菌和弗氏柠檬酸杆菌，进行筛选和阴性选择步骤，以确保所选适体的种属特异性。在对显示对S的最高结合亲和力的池进行测序后，通过自组装将一种对细菌具有高亲和力的DNA适体序列整合到金纳米颗粒修饰的丝网印刷碳电极（GNPs-SPCE）上，从而将其整合到阻抗传感器中。值得注意的是，这种适体传感器具有高度的选择性，可以成功地检测S。在10分钟内将其稀释至600 CFU 1/mL（相当于30 \muL检测体积中的18 CFU），并将其与其他盐藻属物种（包括S.鼠伤寒沙门氏菌（S.猪霍乱该报告设想为使用快速、经济、无标记的电化学平台对各种微生物进行基于适体的分型开辟一个新的场所。
英文摘要：
The development of an aptamer-based impedimetric sensor for typing of bacteria (AIST-B) is presented. Highly specific DNA aptamers to Salmonella enteritidis were selected via Cell-SELEX technique. Twelve rounds of selection were performed; each comprises a positive selection step against S. enteritidis and a negative selection step against a mixture of related pathogens, including Salmonella typhimurium, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Citrobacter freundii to ensure the species-specificity of the selected aptamers. After sequencing of the pool showing the highest binding affinity to S. enteritidis, a DNA aptamer sequence of high affinity to the bacteria was integrated into an impedimetric sensor via self-assembly onto a gold nanoparticles-modified screen-printed carbon electrode (GNPs-SPCE). Remarkably, this aptasensor is highly selective and can successfully detect S. enteritidis down to 600 CFU 1/mL (equivalent to 18 CFU in 30 \muL assay volume) in 10 minutes and distinguish it from other salmonella species, including S. typhimurium and S. choleraesuis. This report is envisaged to open a new venue for the aptamer-based typing of a variety of microorganisms using a rapid, economic, label-free electrochemical platform.
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