Rapid Multiplex Detection of Viral Pathogens in Whole Blood Using Microfluidic Sample Processor and Smartphone-Linked Handheld Instrument

An integrated system for rapid multiplex of whole blood samples for simultaneous detection of Zika, Dengue (serotypes 1 and 3), and Chikungunya is demonstrated using a smartphone assisted detection device and microfluidic sample processor. The microfluidic processor stores all required reagents for sample lysis as well as LAMP master mix and partitions the final solution into 6 compartments for detection of 4 virus assays plus two experimental controls. With a handheld detection instrument that provides uniform 65°C heating, LED illumination, and a smartphone camera for image gathering, the multiplex assay is detected through LAMP-generated fluorescence within the distinct compartments. This system achieves detection limits of live virus spiked into whole blood through the ranges of 2×10<sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">5</sup> - 6×10<sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">6</sup> PFU/ml and lacks cross reactivity between assays while providing a pipette free operation with no additional equipment.