(441g) Microfluidic Chips for Multiplexed Viral DNA Detection

Molecular beacons are a highly sensitive means of detection of DNA sequences with specificity down to the single base pair. To date, this detection concept has not been used in high throughput screening, due to the expense associated with these reagents. Microfluidic devices, however, use mere nano- or picoliter volumes of sample, thus overcoming the cost issue and opening up the use of these highly sensitive molecular beacons for high throughput applications.

Here we present the use of a microfluidic chip to create a combinatorial array of multiplexed molecular beacons against a variance of oligomer targets. This chip utilizes Actuate-to-Open valves which allow for easy filling and isolation of the compartments [1]. Furthermore, the valves do not need to be actuated continuously after filling, so the chip can be disconnected from the ancillary pressure source and transported for further use or analysis.

Biotinylated molecular beacons with different probe regions are specifically bound to a streptavidin-patterned surface in different compartments. Total internal reflection fluorescence (TIRF) will be used to detect whether or not certain viral targets are present across an array of beacons. The key advantage of TIRF in combination with molecular beacons is that the presence of very low concentrations of oligomer target can be detected. This has potential application in point-of-care testing in clinical settings.