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Colorimetric detection of arboviruses and vector mosquito species. (A) Aptamer-gold nanoparticle conjugates, (B) mosquito salivary protein, (C) Zika envelope protein, (D) visible color change, (E) imbibed diagnostic and (F) diagnostic cards.

A polydimethylsiloxane-based microfluidic device has been developed for the multiplex detection of viral envelope proteins such as Zika and chikungunya on a single platform using aptamer–analyte interactions. The channel is integrated with microsized pillars that increase the surface area allowing more aptamers to attach to the incoming envelope protein molecules, thus increasing the overall sensitivity of the system. The working of the device depends on the formation of protein-mediated sandwich morphology that is obtained using an aptamer and aptamer-functionalized gold nanoparticle (AuNP) pair. The colorimetric signal is obtained upon introduction of silver reagents into the channel, which are selectively deposited on the AuNP surface, providing a gray contrast in the testing zone. The microfluidic channel approach successfully detected clinically relevant concentrations of Zika and chikungunya envelope proteins in phosphine-buffered saline (1 pM) and calf blood (100 pM) with high specificity using gold-decorated aptamers integrated in a microfluidic channel.

The rapid spread of viral infections demands early detection strategies to minimize proliferation of the disease. Here, we demonstrate a plasmonic biosensor to detect Dengue virus, which was chosen as a model, via its nonstructural protein NS1 biomarker. The sensor is functionalized with a synthetic single-stranded DNA oligonucleotide and provides high affinity toward NS1 protein present in the virus genome. We demonstrate the detection of NS1 protein at a concentration of 0.1−10 μg/mL in bovine blood using an on-chip microfluidic plasma separator integrated with the plasmonic sensor which covers the clinical threshold of 0.6 μg/mL of high risk of developing Dengue hemorrhagic fever. The conceptual and practical demonstration shows the translation feasibility of these microfluidic optical biosensors for early detection of a wide range of viral infections, providing a rapid clinical diagnosis of infectious diseases directly from minimally processed biological samples at point of care locations.

Morphological modification of gold nanoparticles to obtain a highly efficient paper based sensor for colorimetric detection of epinephrine (LOD – Limit of Detection).

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