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Integrated nucleic acid assays

  • US 8,772,017 B2
  • Filed: 06/08/2012
  • Issued: 07/08/2014
  • Est. Priority Date: 03/15/2006
  • Status: Active Grant
First Claim
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1. A microfluidic cartridge for extracting a nucleic acid fraction from a sample, comprising:

  • a) a loading port for introduction of the sample;

    b) a waste chamber;

    c) a lysis sub-circuit operably linked to said loading port, wherein said cell lysis sub-circuit comprises;

         1) a mixing chamber;

         2) a sealed pouch containing lysis reagent; and

         3) a pump member configured for pumping said lysis reagent into said mixing chamber, wherein said lysis sub-circuit is configured to generate a lysed sample from the sample in said mixing chamber; and

    d) a nucleic acid extraction sub-circuit operably linked to both of said lysis sub-circuit and said waste chamber, wherein said nucleic acid extraction sub-circuit comprises;

         1) a solid phase nucleic acid extraction component configured to bind nucleic acids present in said lysed sample;

         2) a sealed pouch containing wash reagent;

         3) a sealed pouch containing elution reagent; and

         4) a pump member configured for pumping said elution reagent over said solid phase nucleic acid extraction component to generate a mixture of extracted nucleic acids;

    wherein said microfluidic cartridge is configured to pump said lysis reagent through a valved connection into said mixing chamber of said lysis subcircuit and said microfluidic cartridge is configured to pump said elution reagent through a valved connection to said solid phase nucleic acid extraction component of said nucleic acid extraction subcircuit;

    wherein said pump members are configured to be pneumatically driven by applied positive pressure strokes and suction pressure strokes;

    wherein said applied positive pressure strokes and suction pressure strokes enable reciprocating flow through said valved connections; and

    wherein the pump member for pumping said elution reagent is a pneumatically actuatable, elastomeric diaphragm configured to enable slug elution of said mixture of extracted nucleic acids.

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