Abstract
Dengue is an arbovirus disease transmitted mainly by Aedes mosquitoes. As dengue shares similar clinical symptoms with other infectious diseases, prompt and accurate diagnosis is pivotal to clinicians’ decisions on appropriate management. Conventional diagnostic tests to detect the dengue-specific IgM antibody are limited in their performance and ease of use. To address these issues, we developed and evaluated a biosensor based on screen-printed carbon electrodes (SPCEs) for the detection of dengue-specific immunoglobulin M (IgM) antibodies. Various optimisations were performed in order to increase the sensitivity and specificity of the biosensor. For optimal and proper orientation of the paratope sites of goat anti-human IgM capture antibodies (GAHICA), various antibody techniques, including passive, covalent, protein A, protein G and streptavidin/biotin systems, were tested on the SPCEs. The assay reagents for the biosensor were also optimised prior to its evaluation. Analytical sensitivity evaluation was carried out using pooled sera, while analytical specificity evaluation was conducted on a panel of six non-dengue serum samples. Subsequently, diagnostic sensitivity and specificity evaluation were performed using 144 reference samples. Electrochemical current signals generated from H2O2 catalysed by HRP-labelled anti-dengue detection antibodies were measured using the chronoamperometric technique. With a limit of detection (LOD) of 106 serum dilution, the analytical sensitivity of the developed biosensor was 10 times higher than commercial ELISA. The analytical specificity of this dengue IgM biosensor was 100%. Similarly, the biosensor’s diagnostic performance was 100% for sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). These findings suggest that the developed biosensor has a great potential to be used to diagnose dengue after seroconversion.
Highlights
Dengue is among the most important human viral diseases that exert a considerable global health burden
The surface of carbon working electrode (CWE) with immobilised goat anti-human IgM capture antibodies (GAHICA) showed cloudy clusters when compared to blank CWE
An electrochemical biosensor based on the MAC-enzyme-linked immunosorbent assay (ELISA) principle was successfully developed on screen printed carbon electrodes for the detection of dengue immunoglobulin M (IgM) antibodies
Summary
Dengue is among the most important human viral diseases that exert a considerable global health burden. In the past few decades, the global prevalence of dengue has risen significantly. Dengue, which poses risk to approximately 3.6 billion people, is endemic in over 100 tropical and subtropical countries. 390 million dengue cases are estimated to occur. Dengue patients often experience a variety of clinical manifestations, ranging from mild dengue fever to life-threatening severe dengue [1,2]. A vaccine is commercially available, it is not widely accessible in many countries [3]
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