Systems and methods for multiplexed electrochemical detection

• US 9,804,120 B2
• Filed: 11/30/2015
• Issued: 10/31/2017
• Est. Priority Date: 05/24/2012
• Status: Active Grant

• Current Assignee: The Governing Council of the University of Toronto (University of Toronto)
• Original Assignee: The Governing Council of the University of Toronto (University of Toronto)
• Inventors: Kelley, Shana O., Sargent, Edward Hartley, Lam, Brian
• Primary Examiner(s): Bhat, Narayan
• Application Number:
  US14/953,577
• Publication Number:
  US 20160077046A1
• Time in Patent Office: 701 Days
• Field of Search: 435 61, 435 71, 4352831, 4352872, 4352884, 536 243, 422 681, 422 8201, 204400, 204406, 204411
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 2563/137 Metal/ion, e.g. metal label
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode
• CPC Class Codes: G01N 27/27 Association of two or more measuring systems or cells, each measuring a different parameter, where the measurement results may be either used independently, the systems or cells being physically associated, or combined to produce a value for a further parameter
• CPC Class Codes: G01N 27/3275 Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
• CPC Class Codes: G01N 27/403 Cells and electrode assemblies
• CPC Class Codes: G01N 27/416 Systems G01N27/27 takes precedence
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

Contemplated methods and devices comprise performing electrochemical sample analysis in a multiplexed electrochemical detector having reduced electrical cross-talk. The electrochemical detector includes electrodes that share a common lead from a plurality of leads. The sample, which may be a liquid sample, is introduced into one or more sample wells and a signal is applied to at least one of the electrodes. A response signal is measured while simultaneously applying a substantially fixed potential to each of a remainder of the plurality of leads.

Claims

1. A method for performing electrochemical analysis, the method comprising:

introducing a sample into a multiplexed electrochemical detector, the multiplexed electrochemical detector comprising a plurality of leads adapted to be electrically coupled to a first plurality of working electrodes, and a first counter electrode adapted to be electrically coupled to a second plurality of working electrodes, each of the first plurality of working electrodes housed in a different one of a plurality of sample wells, each of the second plurality of working electrodes housed in a first sample well of the plurality of sample wells;

applying a first signal to a first lead of the plurality of leads;

measuring a first response signal from the first counter electrode while simultaneous applying a substantially fixed potential to each of a first remainder of the plurality of leads; and

determining whether a first target analyte is present in the sample based on the first response signal.

2. The method of claim 1, further comprising:

applying a second signal to a second lead of the plurality of leads;

measuring a second response signal from a second counter electrode while simultaneously applying a substantially fixed potential to each of a second remainder of the plurality of leads; and

determining whether a second target analyte is present in the sample based on the second response signal.

3. The method of claim 2, wherein each of the plurality of leads is arranged in one of a plurality of rows on a solid support, and each of the first and second sample wells is formed from first and second elongated channels, respectively, that run substantially perpendicular to each of the plurality of rows.

4. The method of claim 3, wherein each of the first and second channels comprises hydrophobic and hydrophilic regions.

5. The method of claim 2, wherein the electrochemical detector further comprises a first plurality of probes each tethered to one of the first plurality of working electrodes, wherein the first plurality of probes is selected to hybridize with the first target analyte.

6. The method of claim 5, wherein the electrochemical detector further comprises a second plurality of probes tethered to one of the second plurality of working electrodes, wherein the second plurality of probes is selected to hybridize with the second target analyte but not the first target analyte.

7. The method of claim 6, wherein the first and second pluralities of probes are selected from nucleic acids, peptide nucleic acids, locked nucleic acids, proteins, and peptides.

8. The method of claim 2, wherein each of the first and second counter electrodes is coupled to a common potentiostat, and wherein measuring the first and second response signals comprises sequentially measuring the first and second response signals using the common potentiostat.

9. The method of claim 2, wherein the first counter electrode is coupled to a first potentiostat and second counter electrode is coupled to a second potentiostat, the method further comprising sequentially measuring the first and second response signals using the first and second potentiostats, respectively.

10. The method of claim 2, wherein the first and second pluralities of working electrodes comprise one or more nanostructured microelectrodes.

11. The method of claim 1, wherein the first plurality of working electrodes are adapted to be electrically coupled to the first counter electrode by a first fluid portion of the sample that simultaneously contacts each of the first plurality of working electrodes and the first counter electrode.

12. The method of claim 11, wherein the second plurality of working electrodes are adapted to be electrically coupled to the second counter electrode by a second fluid portion of the sample that simultaneously contacts each of the second plurality of working electrodes and the second counter electrode, wherein the second fluid portion is substantially physically isolated from the first fluid portion during said measuring of the first and second response signals.

13. The method of claim 1, further comprising applying to the electrochemical detector an electrochemical reagent comprising Ru(NH₃)₆³⁺ and Fe(CN)₆³⁻

.

14. The method of claim 1, wherein determining whether the first target analyte is present in the sample comprises determining that a magnitude of the first response signal is greater than a threshold value.

15. The method of claim 1, further comprising providing an indicator of whether the first target analyte is present in the sample,

the indicator selected from a magnitude of the first response signal, a concentration of the first target analyte determined based on the first response signal, a color-coded indicator selected based on the first response signal, a symbol selected based on the first response signal, a graphical representation of the first response signal over a plurality of values of the first voltage signal, and any suitable combination thereof.

16. The method of claim 1, wherein the sample is a liquid sample.

17. The method of claim 1, wherein the sample contains bacteria.

18. The method of claim 17, wherein the bacteria in the sample are lysed prior to introducing the sample into the multiplexed electrochemical detector.

19. The method of claim 1, wherein the substantially fixed potential is selected to reduce current flow from the first remainder of the plurality of leads to the first counter electrode.

20. The method of claim 19, wherein the substantially fixed potential corresponds to a ground potential.

21. The method of claim 1, wherein each of the plurality of leads extend in a first direction and the first counter electrode extends in a second direction different from the first direction.

22. The method of claim 1, wherein the second plurality of working electrodes includes at least one of the first plurality of working electrodes.

23. The method of claim 1, wherein the multiplexed electrochemical detector further comprises a reference electrode.

24. The method of claim 23, wherein the reference electrode is adapted to be electrically coupled to the second plurality of working electrodes.