Integrated nucleic acid assays
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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Abstract
Integrated microfluidic cartridges for nucleic acid extraction, amplification, and detection from clinical samples are disclosed. The devices are single-entry, sanitary, and disposable. The devices enable simplex or multiplex nucleic acid target detection, as for example: assay panels for multiple infectious agents, or assay panels for cancerous cell types. Methods for use of microfluidic cartridges in a fully automated, pneumatically controlled apparatus are also disclosed.
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Citations
10 Claims
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1. A microfluidic cartridge for extracting a nucleic acid fraction from a sample, comprising:
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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; andd) 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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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Specification