A microfluidic cartridge assembly for performing an assay of a target analyte in a biological sample, the microfluidic cartridge comprising: a cartridge housing defining one or more reservoirs, the one or more reservoirs including a reagent reservoir containing one or more fluid reagents; a microfluidic substrate attached to the cartridge housing, the microfluidic substrate having a plurality of microfluidic channels in fluid communication with the one or more reservoirs, the plurality of microfluidic channels being adapted for performing an assay of the target analyte in the sample using the one or more fluid reagents; one or more valves in fluid communication with the plurality of microfluidic channels that are actuatable between a closed configuration that inhibits fluid flow through the microfluidic channels and an open configuration allowing fluid flow from the one or more reservoirs through the plurality of microfluidic channels, wherein the one or more valves include a first valve positioned such that, when in the closed configuration, the first valve isolates the one or more fluid reagents in the reagent reservoir from the plurality of microfluidic channels and, when in the open configuration, the first valve allows flow of the one or more fluid reagents through the microfluidic channels; and one or more pressure septa in fluid communication with the plurality of microfluidic channels adapted for introducing pressure into the plurality of microfluidic channels such that introducing a pressure into the plurality of microfluidic channels allows fluid flow through the plurality of microfluidic channels in a controlled manner so as to perform the assay of the target analyte in the sample using the one or more fluid reagents during fluid flow through the plurality of microfluidic channels when the one or more valves are in the open configuration. a cartridge housing defining one or more reservoirs, the one or more reservoirs including a reagent reservoir containing one or more fluid reagents; a microfluidic substrate attached to the cartridge housing, the microfluidic substrate having a plurality of microfluidic channels in fluid communication with the one or more reservoirs, the plurality of microfluidic channels being adapted for performing an assay of the target analyte in the sample using the one or more fluid reagents; one or more valves in fluid communication with the plurality of microfluidic channels that are actuatable between a closed configuration that inhibits fluid flow through the microfluidic channels and an open configuration allowing fluid flow from the one or more reservoirs through the plurality of microfluidic channels, wherein the one or more valves include a first valve positioned such that, when in the closed configuration, the first valve isolates the one or more fluid reagents in the reagent reservoir from the plurality of microfluidic channels and, when in the open configuration, the first valve allows flow of the one or more fluid reagents through the microfluidic channels; and one or more pressure septa in fluid communication with the plurality of microfluidic channels adapted for introducing pressure into the plurality of microfluidic channels such that introducing a pressure into the plurality of microfluidic channels allows fluid flow through the plurality of microfluidic channels in a controlled manner so as to perform the assay of the target analyte in the sample using the one or more fluid reagents during fluid flow through the plurality of microfluidic channels when the one or more valves are in the open configuration.