Assay cartridges and methods of using the same

A method of conducting a PCR analysis of a sample in an assay cartridge, said cartridge comprising a fluidic network and a plurality of chambers, wherein said fluidic network comprises:

• (a) a primary flow path comprising, from a proximate to a distal end, (i) an inlet, (ii) a purification zone, (iii) a PCR reaction zone including a first reaction temperature controlled zone and a second reaction temperature controlled zone, and (iv) a detection zone, wherein said primary flow path further comprises (vi) one or more air vent ports, and
• (b) one or more fluidic conduits each intersecting said primary flow path and fluidically connected to one or more of said plurality of chambers, wherein each of said plurality of chambers are connected to an additional air vent port,
• wherein said plurality of chambers include:
• a sample chamber, a lysis reagent chamber, a lysis chamber, a purification reagent chamber, a plurality of PCR reagent chambers, and one or more waste chambers;
• said method comprising the steps of:
• (i) interfacing the fluidic network with a fluidic control manifold configured to drive fluid motion within said fluidic network.
• (ii) metering a volume of sample from said sample chamber to said lysis chamber;
• (iii) metering a volume of lysis buffer from said lysis reagent chamber to said lysis chamber;
• (iv) lysing said volume of sample;
• (v) moving lysate from said lysis chamber to said purification zone;
• (vi) extracting nucleic acid from said lysate;
• (vii) purifying said nucleic acid;
• (viii) moving said a purified nucleic acid to said PCR reaction zone;
• (ix) contacting said purified nucleic acid with one or more PCR reagents to form a mixture;
• (x) shuttling said mixture formed in step (ix) between said first and second temperature controlled zones;
• (xi) repeating steps (ix) and (x) to form an amplified product mixture;
• (xii) contacting said amplified product mixture with a detection reagent;
• (xiii) moving said mixture formed in step (xi) to said detection zone;
• and
• (xiv) measuring a signal from said detection zone.