Abstract
Early diagnosis of dengue biomarkers by employing a technology that is less labor- and time-intensive and offers higher sensitivity and lower limits of detection would find great significance in the developing world. Here, we report the development of a biosensor that exploits the localized surface plasmon resonance (LSPR) effect of silver nanostructures, created via thermal annealing of thin metal film, to detect dengue NS1 antigen, which appears as early as the onset of infection. The biosensor integrates membrane-based blood-plasma separation to develop lab-on-chip device that facilitates rapid diagnosis (within 30 min) of dengue NS1 antigen from a small volume (10 µL) of whole blood. The refractive index (RI) sensitivity of the LSPR biosensor was verified by using aqueous glycerol (0-100 wt%) which showed that it is sufficiently sensitive to detect 10-3 change in RI, which is comparable to that observed with protein-protein interaction. The RI sensitivity was utilized to demonstrate protein binding by using bovine serum albumin and detection of antibody-antigen immune reaction by binding human chorionic gonadotropin antigen to immunoglobulin antibody immobilized in our LSPR biosensor. Next, we demonstrated the detection of NS1 in plasma obtained via centrifugation and in plasma separated on-chip. From 10 µL of whole blood spiked with NS1 antigen, our biosensor reliably detects 0.06 µg/mL of NS1, which lies within the clinical limit observed during the first seven days of infection, with a sensitivity of 9 nm/(µg/mL). These results confirm that the proposed LSPR biosensor can potentially be used in point-of-care dengue diagnostics.