Screen Printed Carbon Electrode Based Electrochemical Immunosensor for the Detection of Dengue NS1 Antigen

Diagnostics

Om Parkash²  Chan Yean Yean²  Rafidah Hanim Shueb¹ 

[1] Department of Medical Microbiology and Parasitology, School of Medical Science, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia;

关键词: electrochemical immunosensor;    dengue diagnosis;    NS1;    screen printed carbon electrode;    streptavidin/biotin;   

DOI  :  10.3390/diagnostics4040165

来源: mdpi

【 摘 要 】

An electrochemical immunosensor modified with the streptavidin/biotin system on screen printed carbon electrodes (SPCEs) for the detection of the dengue NS1 antigen was developed in this study. Monoclonal anti-NS1 capture antibody was immobilized on streptavidin-modified SPCEs to increase the sensitivity of the assay. Subsequently, a direct sandwich enzyme linked immunosorbent assay (ELISA) format was developed and optimized. An anti-NS1 detection antibody conjugated with horseradish peroxidase enzyme (HRP) and 3,3,5,5'-tetramethybezidine dihydrochloride (TMB/H₂O₂) was used as an enzyme mediator. Electrochemical detection was conducted using the chronoamperometric technique, and electrochemical responses were generated at −200 mV reduction potential. The calibration curve of the immunosensor showed a linear response between 0.5 µg/mL and 2 µg/mL and a detection limit of 0.03 µg/mL. Incorporation of a streptavidin/biotin system resulted in a well-oriented antibody immobilization of the capture antibody and consequently enhanced the sensitivity of the assay. In conclusion, this immunosensor is a promising technology for the rapid and convenient detection of acute dengue infection in real serum samples.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.