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Development of Electrochemical Impedance Spectroscopy Based Malaria Aptasensor Using HRP‐II as Target Biomarker
Electroanalysis ( IF 2.7 ) Pub Date : 2018-04-25 , DOI: 10.1002/elan.201800142 Babina Chakma 1 , Priyamvada Jain 1 , Naveen Kumar Singh 1 , Pranab Goswami 1
Electroanalysis ( IF 2.7 ) Pub Date : 2018-04-25 , DOI: 10.1002/elan.201800142 Babina Chakma 1 , Priyamvada Jain 1 , Naveen Kumar Singh 1 , Pranab Goswami 1
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Current demand for a stable, low cost and sensitive malaria sensor has prompted to explore novel recognition systems that can substitute widely used protein based labile biorecognition elements to be used in point of care diagnostic devices. Here, we report a novel ssDNA aptamer of 90 mer sequence developed by SELEX process against HRP‐II, a specific biomarker for Plasmodium falciparum strains. High stability of the secondary structure of the isolated aptamer was discerned from its free energy of folding of −20.40 kcal mole−1. The binding constant (Kd) of the aptamer with HRP‐II analysed by isothermal titration calorimetry was ∼1.32 μM. Circular dichroism studies indicated B form of the aptamer DNA. The aptamer was chemically immobilized on a gold electrode surface through a self‐assembled monolayer of dithio‐bis(succinimidyl) propionate to produce the aptasensor. The step wise modification of the layers over the gold electrode during fabrication of the aptasensor was confirmed by cyclic voltammetry. The aptasensor was then challenged with different concentration of HRP‐II and analysed the interaction signals through electrochemical impedance spectroscopy. The impedance signal behaved reciprocally with the increasing concentrations of the target in the sample from which a dynamic range of 1 pM–500 pM (R2=0.99) and LOD of ∼3.15 pM were discerned. The applicability of the developed aptasensor to detect HRP‐II in mimicked real sample was also validated.
中文翻译：
基于HRP-II作为目标生物标记物的基于电化学阻抗谱的疟疾适应传感器的开发
当前对稳定，低成本和灵敏的疟疾传感器的需求促使人们探索新颖的识别系统，该系统可以替代广泛使用的基于蛋白质的不稳定生物识别元件，以用于护理点诊断设备。在这里，我们报告了一种新型的90 mer序列的ssDNA适体，它是通过SELEX程序针对HRP-II（恶性疟原虫菌株的一种特定生物标记）开发的。分离的适体的二级结构的高稳定性从其-20.40kcal mole -1的折叠自由能中看出。结合常数（K d用HRP-II等温滴定热分析法分析的适体约为1.32μM。圆二色性研究表明适体DNA为B型。适体通过二硫代双（琥珀酰亚胺基）丙酸酯的自组装单层化学固定在金电极表面上，以产生适体传感器。通过循环伏安法证实了在适体传感器的制造过程中金电极上的层的逐步修饰。然后用不同浓度的HRP-II挑战aptasensor，并通过电化学阻抗谱分析相互作用信号。阻抗信号与样品中靶标浓度的增加成反比，其动态范围为1 pM–500 pM（R 2= 0.99）且LOD为〜3.15 pM。还验证了开发的适体传感器在模拟真实样品中检测HRP-II的适用性。
更新日期：2018-04-25
中文翻译：
基于HRP-II作为目标生物标记物的基于电化学阻抗谱的疟疾适应传感器的开发
当前对稳定，低成本和灵敏的疟疾传感器的需求促使人们探索新颖的识别系统，该系统可以替代广泛使用的基于蛋白质的不稳定生物识别元件，以用于护理点诊断设备。在这里，我们报告了一种新型的90 mer序列的ssDNA适体，它是通过SELEX程序针对HRP-II（恶性疟原虫菌株的一种特定生物标记）开发的。分离的适体的二级结构的高稳定性从其-20.40kcal mole -1的折叠自由能中看出。结合常数（K d用HRP-II等温滴定热分析法分析的适体约为1.32μM。圆二色性研究表明适体DNA为B型。适体通过二硫代双（琥珀酰亚胺基）丙酸酯的自组装单层化学固定在金电极表面上，以产生适体传感器。通过循环伏安法证实了在适体传感器的制造过程中金电极上的层的逐步修饰。然后用不同浓度的HRP-II挑战aptasensor，并通过电化学阻抗谱分析相互作用信号。阻抗信号与样品中靶标浓度的增加成反比，其动态范围为1 pM–500 pM（R 2= 0.99）且LOD为〜3.15 pM。还验证了开发的适体传感器在模拟真实样品中检测HRP-II的适用性。
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