Abstract
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.
Highlights
Dengue continues to be a major public health concern in tropical and subtropical countries
The screen printed carbon electrodes (SPCEs) were subjected to 10 cycles of cyclic voltammetry (CV) at 100 mV/s scan rate in 5 mM ferricyanide solution prepared in 0.1 M KCl
We have proposed a simple approach to the development of an electrochemical immunosensor for the detection of the dengue nonstructural 1 (NS1) antigen in real samples
Summary
Dengue continues to be a major public health concern in tropical and subtropical countries. There are over 3.6 billion people at risk from dengue infection. It is estimated that 390 million dengue infections occur worldwide [1,2]. This leads to 250,000–500,000 cases of dengue hemorrhagic fever (DHF) with 5%–10% mortality annually [3,4]. Early and prompt diagnosis of dengue can help patient triage and care management [5,6]. Routine laboratory methods used for the diagnosis of dengue infection are viral culture, viral nucleic acid amplification by reverse transcriptase PCR (RT-PCR), and serological tests like immunoglobulin M (IgM) capture ELISA and immunoglobulin G (IgG) capture
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