Methods and devices for detecting SARS-COV-2

A method for detecting the presence of a target-nucleic acid sequence in a nucleic acid-containing sample comprising:

• a) receiving an electrolytic fluid solution including ions and a nucleic acid-containing sample in a fluid chamber having a first and second electrode, wherein at least one electrode has attached thereto a plurality of capture/primer-probes complementary to the target-nucleic acid, wherein the capture/primer-probes are at the electrode-solution interface;
• b) applying an annealing-voltage to the electrolytic fluid in proximity to the electrode-solution interface to generate an annealing-pH level by the interface, wherein the annealing-pH level causes annealing of any number of complementary base pairs between nucleic acid within the nucleic acid-containing sample and the capture/primer-probes;
• c) applying a denaturing-voltage to the electrolytic fluid in proximity to the electrode-solution interface to generate a denaturing-pH level by the interface, wherein the denaturing-pH level causes the denaturation of double-stranded nucleic acids, including hybridized pairs of capture/primer-probes and target-nucleic acids, that are either a particular number of base pairs shorter or a particular range of base pairs shorter than the full-length of a double-stranded capture-probe;
• and
• d) modulating the annealing-voltage and denaturing-voltage, wherein multiple cycles of modulation are performed on the entire nucleic acid-containing sample while it is inside the fluid chamber, and wherein after the final application of the corresponding pH level, the target-nucleic acid is enriched or isolated from the nucleic acid-containing sample by remaining bound to the capture/primer-probe on the at least one electrode, wherein the bound target-nucleic acid is a template strand for template directed elongation synthesis, wherein the modulating voltages between the annealing-voltage and denaturing-voltage is at a modulation frequency in the range of 0.1-1000 Khz;
• e) providing an elongation mixture comprising (i) a polymerase enzyme, (ii) an ATP regenerating enzyme, (iii) a luminescence enzyme, and (iv) a polymerase-ATP regenerating enzyme-luminescence reagent solution having the components for carrying out template directed elongation synthesis of a growing nucleic acid strand, wherein said reagent solution includes an ATP-regenerating-enzyme-substrate, a luminescence-substrate;
• and a plurality of types of dNTPs or nucleotide analogs, wherein each type of nucleotide analog has a leaving group that is cleavable by the polymerase, wherein the leaving group is cleaved upon polymerase-dependent binding of a respective nucleotide analog to the template strand;
• f) carrying out nucleic acid elongation synthesis such that one or a plurality of nucleotide analogs are added sequentially to the template strand if the capture/primer-probe hybridizes to the target nucleic acid sequence, whereby:
• a) a nucleotide analog associates with the polymerase, b) the nucleotide analog is incorporated on the template strand by the polymerase when the leaving group on that nucleotide analog is cleaved by the polymerase, wherein the leaving group is combined with an ATP-regenerating-enzyme-substrate by the ATP regenerating enzyme yielding ATP, then c) binding the ATP to a luminescence-enzyme, wherein a luminescence-substrate is catalyzed by the luminescence-enzyme to produce luminescence while regenerating the respective leaving group;
• and
• g) detecting light from the luminescence while nucleic acid synthesis is occurring, whereby detection of light indicates the presence of the particular target nucleic acid sequence.