Systems and methods for multiplexed electrochemical detection

• US 9,217,179 B2
• Filed: 03/15/2013
• Issued: 12/22/2015
• 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 H., Lam, Brian
• Primary Examiner(s): Bhat, Narayan
• Application Number:
  US13/843,323
• Publication Number:
  US 20130316340A1
• Time in Patent Office: 1,012 Days
• Field of Search: 435/6.1, 435/283.1, 435/288.4, 422/68.1, 422/82.01, 204/400, 204/406, 204/411
• US Class Current: 1/1
• 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 multiplexed electrochemical detection system, comprising:

a solid support;

a plurality of leads affixed to the solid support, each of the plurality of leads extending in a first direction in the solid support and being adapted to be electrically coupled to a first plurality of working electrodes, wherein each of the first plurality of working electrodes is housed in a different one of a plurality of sample wells within the solid support;

a first counter electrode extending in a second direction different from the first direction and adapted to be electrically coupled to a second plurality of working electrodes, wherein the second plurality of working electrodes includes at least one of the first plurality of working electrodes, and wherein each of the second plurality of working electrodes is housed in a first sample well in the plurality of sample wells;

a first signal input interface adapted to be electrically coupled to a first lead of the plurality of leads and adapted to selectively apply a first signal to the first lead;

a second signal input interface adapted to be electrically coupled to a first subset of the plurality of leads and adapted to selectively apply a substantially fixed potential to each of the first subset of the plurality of leads; and

a detection circuitry configured to;

measure a first response signal from the first counter electrode while the second signal input interface applies the substantially fixed potential to the remainder of the second plurality of leads.

2. The system of claim 1, further comprising:

a second counter electrode adapted to be electrically coupled to a third plurality of working electrodes including at least one of the first plurality of working electrodes, wherein;

the first signal input interface is adapted to be electrically coupled to a second lead of the plurality of leads and adapted to selectively apply a third signal to the second lead;

the second signal input interface adapted to be electrically coupled to a second subset of the plurality of leads and adapted to selectively apply a substantially fixed potential to each of the second subset of the plurality of leads, and

the detection circuitry is further configured to;

measure a second response signal from the second counter electrode while while the second signal input interface applies the substantially fixed potential.

3. The system of claim 2, wherein the second plurality of working electrodes are adapted to be electrically coupled to the first counter electrode by the presence of a first fluid portion of the sample capable of simultaneously contacting each of the second plurality of working electrodes and the first counter electrode.

4. The system of claim 3, wherein the third plurality of working electrodes are adapted to be electrically coupled to the second counter electrode by the presence of a second fluid portion of the sample capable of simultaneously contacting each of the third 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.

5. The system of claim 4, wherein each of the plurality of leads is arranged in one of a plurality of rows on the solid support, and each of the first and second sample wells is formed from first and second elongated channels, respectively, that run perpendicular to each of the plurality of rows.

6. The system of claim 5, wherein each of the first and second channels comprise hydrophobic and hydrophilic regions.

7. The system of claim 6, further comprising a first plurality of probes tethered to at least some of the second plurality of working electrodes, wherein the first plurality of probes is selected to hybridize with a first target analyte.

8. The system of claim 7, further comprising a second plurality of probes tethered to at least some of the third plurality of working electrodes, wherein the second plurality of probes is selected to hybridize with a second target analyte but not the first target analyte.

9. The system of claim 8, wherein each of the first and second counter electrodes are adapted to couple to a common potentiostat, and wherein the detection circuitry is further configured to measure the first and second response signals sequentially using the common potentiostat.

10. The system of claim 8, wherein the first counter electrode is adapted to couple to a first potentiostat and second counter electrode is adapted to couple to a second potentiostat, and wherein the detection circuitry is further configured to measure the first and second response signals sequentially using the first and second potentiostats, respectively.

11. The system of claim 8, wherein the first and second pluralities of working electrodes comprise one or more nanostructured microelectrodes.

12. The system of claim 11, further comprising applying to the electrochemical detector an electrochemical reagent comprising Ru(NH3)₆³⁺ and Fe(CN)₆³⁻

.

13. The system of claim 12, wherein the first and second pluralities of probes are selected from the group consisting of nucleic acids, peptide nucleic acids, locked nucleic acids, proteins, and peptides.

14. The system of claim 13, wherein the detection circuitry is configured to interface with a processor, the processor being configured to determine that a magnitude of the first response signal is greater than a threshold value.

15. The system of claim 14, wherein the processor is further configured to provide an indicator of whether the first target analyte is present in the sample, the indicator selected from the group consisting of amplitude 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 system of claim 15, further comprising a plurality of reference electrodes, each adapted to be electrically coupled to one of the plurality of sample wells, the plurality of reference electrodes including a first reference electrode adapted to be coupled to the first sample well.

17. The system of claim 16, wherein the first reference electrode is paired with the first counter electrode.

18. The system of claim 17, wherein the sample is a liquid sample.

19. The system of claim 18, wherein the sample contains bacteria.

20. The system of claim 19, further comprising a lysing channel that lyses the bacteria in the sample prior to introducing the sample into the multiplexed electrochemical detector.

21. The system of claim 1, wherein the substantially fixed potential is selected to reduce current flow from the first subset of the plurality of leads to the first counter electrode.

22. The system of claim 1, wherein the substantially fixed potential corresponds to a ground potential.