Methods and apparatus for nanomembrane-based nucleic acid sensing platform for portable diagnostics
A DNA/RNA detection technology is provided. The open flow detection technique includes a substrate defining a pair of opposing microchannels, a pair of opposing electrodes in the opposing microchannels, and at least one ion exchanging nanomembrane coupled between the opposing microchannels such that the opposing microchannels are connected to each other only through the nanomembrane, wherein the nanomembrane is functionalized with a probe complementary to the macromolecule. A voltammeter is provided to measure the electrical current or potential across the nanomembrane, and detect a change in the measured electrical current or potential to quantify the presence of the macromolecule.
1. A quantitative method for determining a target nucleic acid concentration, the method comprising:
making an assay mixture containing the target nucleic acid, wherein the target nucleic acid comprises an electrical charge;
providing a microchamber cell comprising:
a substrate defining a pair of opposing microchannels; a first pair of opposing electrodes in the opposing microchannels for applying a source signal and a second pair of opposing electrodes for measuring an output signal; and
at least one ion exchanging nanomembrane having a charge opposite to that of the target nucleic acid coupled between the opposing microchannels such that the opposing microchannels are connected to each other only through the nanomembrane, wherein the nanomembrane is functionalized with a probe complementary to the target nucleic acid wherein the nanomembrane is provided in a nanoslot;
flowing the assay mixture through one of the microchannels of the microchamber electrochemical cell such that assay mixture contacts the nanomembrane in a manner suitable for hybridization of the assay mixture with the probe, while the other microchannel acts as an auxiliary channel;
connecting the first pair of opposing electrodes to the device to charge the nanomembrane with the source signal;
connecting the second pair of opposing electrodes for measuring the output signal across the nanomembrane; and
detecting a shift to lower resistance in a Warburg impedance spectrum of a nucleic acid solution relative to a control without nucleic acid to determine the concentration of the target nucleic acid.
2. The method of claim 1 , wherein a depth of the nanoslot is substantially the same as a depth of the microchannel.
3. The method of claim 1 , further comprising providing an AC current across the pair of opposing electrodes.
4. The method of claim 1 , wherein the target nucleic acid is at least one of a DNA biomolecule or an RNA biomolecule.
5. The method of claim 1 , further comprising deionizing the assay mixture proximate the nanomembrane.
6. The method of claim 1 , further comprising bulk-to-membrane ion flux over the depletion region formed by the membrane at over-potentials.
7. The method of claim 1 , further comprising forming a non-uniform electroosmotic flow.
8. The method of claim 7 , wherein the non-uniform electroosmotic flow leads to microvorticies formed in the fluid flow.
9. The method of claim 7 , wherein the non-uniform electroosmotic flow leads to a detectable enhanced ion current.
10. The method of claim 1 , further comprising selecting an optimum frequency for each of a particular electrolyte strength wherein the optimum frequency has a sharp minimum in detection time, and wherein the optimum frequency scales as a function of D/λ 2 where D is a molecular diffusivity of the target nucleic acid and λ is the Debye length for the particular electrolyte strength.