Isothermal nucleic acid amplification reactor with integrated solid state membrane
Provided are devices adapted to isolate, amplify, and detect nucleic acids that may be present in a biological sample. The devices can, in some embodiments, isolate, amplify, and detect nucleic acid in a single chamber. In other embodiments, the devices are adapted to isolate and amplify nucleic acids in a reaction chamber, after which the nucleic acids may be communicated to a pervious medium—such as a lateral flow strip—where the user may label and detect the nucleic acids.
1. A method for amplifying nucleic acids, comprising:
contacting a nucleic acid isolation membrane with a nucleic acid under conditions such that the nucleic acid binds to the nucleic acid isolation membrane,
the nucleic acid isolation membrane being disposed in a cassette body; and
effecting relative motion between the cassette body and a cassette holder that engages with the cassette body so as to effect sequential washing of the nucleic acid by a wash fluid stored in the cassette body, suspension of the nucleic acid in a reaction chamber of the cassette body, and amplification of at least some of the nucleic acid.
2. The method of claim 1 , wherein the washing comprises applying a first wash buffer to the nucleic acid isolation membrane.
3. The method of claim 1 , wherein engagement between an actuator of the cassette holder and a wash fluid container of the cassette body effects release of the wash fluid from the wash fluid container.
4. The method of claim 1 , further comprising applying a second wash fluid to the nucleic acid.
5. The method of claim 1 , further comprising detecting the amplified nucleic acid.
6. The method of claim 5 , wherein detection of the amplified nucleic acid is indicative of disease or infection.
7. The method of claim 1 , wherein suspension of the bound nucleic acid comprises suspending the nucleic acid in a suspension fluid.
8. The method of claim 1 , wherein engagement between an actuator of the cassette holder and a suspension fluid container of the cassette body effects release of the fluid from the suspension fluid container so as to suspend the nucleic acid.
9. The method of claim 1 , wherein the relative motion of the cassette body and the cassette holder is effected in an automated fashion.
10. The method of claim 1 , further comprising placing the cassette body into engagement with a heat source.
11. The method of claim 3 , wherein the actuator comprises a projection and the wash fluid container comprises a deformable region that engages with the projection.