Development of a thermochromic lateral flow assay to improve sensitivity for dengue virus serotype 2 NS1 detection

Abstract

Dengue disease is a viral infection that has been widespread in tropical regions, such as Southeast Asia, South Asia and South America. A worldwide effort has been made over a few decades to halt the spread of the disease and reduce fatalities. Lateral flow assay (LFA), a paper-based technology, is used for dengue virus detection and identification because of its simplicity, low cost and fast response. However, the sensitivity of LFA is relatively low and is usually insufficient to meet the minimum requirement for early detection. In this study, we developed a colorimetric thermal sensing LFA format for the detection of dengue virus NS1 using recombinant dengue virus serotype 2 NS1 protein (DENV2-NS1) as a model antigen. Plasmonic gold nanoparticles, including gold nanospheres (AuNSPs) and gold nanorods (AuNRs), and magnetic nanoparticles (MNPs), namely iron oxide nanoparticles (IONPs) and zinc ferrite nanoparticles (ZFNPs), were studied for their thermal properties for sensing assays. AuNSPs with 12 nm diameter were chosen due to their great photothermal effect against light-emitting diodes (LEDs). In the thermal sensing assay, a thermochromic sheet is used as a temperature sensor transforming heat into a visible colour. In the typical LFA, the test line is visible at 6.25 ng mL⁻¹ while our thermal sensing LFA offers a visual signal that can be observed at as low as 1.56 ng mL⁻¹. The colorimetric thermal sensing LFA is capable of reducing the limit of detection (LOD) of DENV2-NS1 by 4 times compared to the typical visual readout. The colorimetric thermal sensing LFA can enhance the sensitivity of detection and deliver visuality to the user to translate without the need for an infrared (IR) camera. It has the potential to expand the utilities of LFA and satisfy early diagnostic applications.