Dutta Reshmi, Thangapandi K, Mondal Sumantra, Nanda Amalesh, Bose Shreyosi, Sanyal Shairee, Jana Saikat Kumar, Ghorai Suvankar
Department of Biotechnology, Faculty of Bioengineering, SRM Institute of Science and Technology, Kattankulathur, Tamil Nadu, India.
Department of Biotechnology, National Institute of Technology, Yupia, Papum Pare, Arunachal Pradesh, India.
Avicenna J Med Biotechnol. 2020 Apr-Jun;12(2):77-84.
Dengue burden is increasing day-by-day globally. A rapid, sensitive, cost-effective early diagnosis kit is the need of the hour. In this study, a label-free electrochemical immunosensor was proposed for dengue virus detection. A modified Polyaniline (PANI) coated Glassy Carbon (GC) electrode, immobilized with DENV NS1 antibody was used to detect the circulating DENV NS1 antigen in both spiked and infected sample.
Cloning, purification and expression of DENV NS1 protein in was performed and sensor design, PANI modification on GC electrode surface by electrochemical polymerization and immobilization of NS1 antibody on the modified electrode surface was done and finally the analytical performance of the electrochemical immunosensor was done using Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS).
CV and EIS were used to study and quantitate the circulating DENV antigen. The calibration curve showed wide linearity, good sensitivity (Slope=13.8% IpR/ ) and distribution of data with a correlation coefficient (R) of 0.997. A lower Limit of Detection (LOD) was found to be 0.33 which encourages the applicability of the sensor.
Thus, a PANI based new electrochemical immunosensor has been developed which has the potential to be further modified for the development of cost effective, point of care dengue diagnostic kit.
全球登革热负担日益加重。当下急需一种快速、灵敏且经济高效的早期诊断试剂盒。在本研究中，提出了一种用于检测登革热病毒的无标记电化学免疫传感器。使用修饰有聚苯胺（PANI）的玻碳（GC）电极，固定登革热病毒非结构蛋白1（DENV NS1）抗体，用于检测加标样本和感染样本中循环的DENV NS1抗原。
进行了DENV NS1蛋白在大肠杆菌中的克隆、纯化和表达，完成了传感器设计、通过电化学聚合对GC电极表面进行PANI修饰以及将NS1抗体固定在修饰电极表面，最后使用循环伏安法（CV）和电化学阻抗谱（EIS）对电化学免疫传感器的分析性能进行了检测。
使用CV和EIS研究并定量循环的DENV抗原。校准曲线显示出宽线性范围、良好的灵敏度（斜率 = 13.8% IpR/）以及数据分布，相关系数（R）为0.997。检测限（LOD）较低，为0.33 ，这促使该传感器具有适用性。
因此，已开发出一种基于PANI的新型电化学免疫传感器，其有潜力进一步改进以开发经济高效的即时检测登革热诊断试剂盒。
