Multiplexed Capillary-Flow Immunoassay And Methods of Use Thereof

A multiplex assay device comprising:

• a microfluidic assembly including:
• a sample inlet;
• microfluidic network forming a plurality of microfluidic pathways each extending to a detection area, wherein each of said plurality of microfluidic pathways is in communication with said sample inlet, and further configured to detect a unique target analyte;
• a first dried reagent disposed along each of said microfluidic pathways;
• and
• a second dried reagent disposed each of said microfluidic pathways;
• wherein, when a fluid is provided to the fluid inlet, the fluid fills the microfluidic network and directs a portion of the fluid through each of the microfluidic pathways to the detection area, and wherein the fluid rehydrates the first dried reagent disposed along each of said microfluidic pathways to produce a first rehydrated reagent, and rehydrates the second dried reagent disposed along each of said microfluidic pathways to produce a second rehydrated reagent, wherein the first rehydrated reagent and the second rehydrated reagent are sequentially delivered through their respective microfluidic pathways to the detection area by capillary-driven flow, and
• wherein, when the fluid flows through the microfluidic network, an air bubble is formed within the network thereby restricting communication between the microfluidic pathways.