Signal detection techniques for the detection of analytes

• US 6,740,518 B1
• Filed: 09/17/1999
• Issued: 05/25/2004
• Est. Priority Date: 09/17/1998
• Status: Expired due to Term

• Current Assignee: Roche Molecular Systems (Roche Holding AG)
• Original Assignee: Clinical Micro Sensors Inc. (Roche Holding AG)
• Inventors: Olsen, Gary T., Gonzalez, Javier, Duong, Hau H., Litvack, Daniel A., Kayyem, Jon Faiz, Terbrueggen, Robert H., OʼConnor, Stephen D.
• Primary Examiner(s): Whisenant, Ethan
• Assistant Examiner(s): LU, FRANK WEI MIN
• Application Number:
  US09/397,957
• Time in Patent Office: 1,712 Days
• Field of Search: 435/6, 435/91.1, 435/91.2, 435/183, 435/283.1, 435/287.1, 435/287.2, 435/7.1, 436/94, 536/23.1, 536/24.3, 536/24.33, 536/25.3, 530/300, 530/350
• US Class Current: 435/287.2
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 27/3277 being a redox reaction, e.g. detection by cyclic voltammetry voltammetry per se G01N27/42, G01N27/48
• CPC Class Codes: G01N 27/416 Systems G01N27/27 takes precedence
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

The invention relates to the use of signal processing methods in order to acheive higher signal to noise ratios, to increase the detection limits of target analytes. These techniques include the monitoring of the output signal at higher harmonic frequencies.

Claims

1. A method of determining the presence of target analytes in a sample comprising:

a) applying said sample to an array comprising a plurality of electrodes, wherein at least one electrode comprises an assay complex comprising;

i) a capture binding ligand covalently attached to said electrode;

ii) a target analyte; and

iii) an electron transfer moiety;

b) applying an input waveform to said electrode to generate an output waveform comprising at least one harmonic component, having a harmonic number greater than or equal to two;

c) detecting said output waveform at said electrode;

d) analyzing said harmonic component with harmonic number greater than or equal to two to determine the presence of said target analytes.

2. A method according to claim 1 wherein said target analyte is a nucleic acid.

3. A method according to claim 1 wherein said target analyte is a protein.

4. A method according to claim 1, wherein said analyzing comprises the use of a peak recognition scheme.

5. A method according to claim 1, wherein said analyzing comprises a digital filter.

6. The method of claim 1, wherein said electrode has an asymmetrical response to said input waveform.

7. The method of claim 6, wherein said electron transfer moiety is degradable.

8. The method of claim 7, wherein said electron transfer moiety is luminol.

9. The method of claim 6, further comprising adding a co-reductant to said sample.

10. The method of claim 9, wherein said co-reductant is ferrocyanide.

11. The method of claim 9, wherein said co-reductant has a lower redox potential than said electron transfer moiety.

12. The method of claim 6, further comprising adding a co-oxidant to said sample.

13. The method of claim 1, wherein said input waveform is a voltage waveform and said output waveform is a current waveform, wherein said input waveform comprises an AC component having a first frequency and a first amplitude, and wherein said first amplitude is selected such that said output waveform comprises at least one non-linear harmonic component.

14. The method of claim 1, wherein said harmonic component is chosen from the group of harmonic components consisting of a second, third, fourth, fifth, sixth, seventh, eighth, ninth, and tenth harmonic components.

15. The method of claim 1, wherein the output waveform comprises a plurality of harmonic components and said method comprises analyzing the plurality of harmonic components.

16. The method of claim 1, wherein said input waveform is a voltage waveform comprising a square wave.

17. The method of claim 16, wherein said harmonic component is an even harmonic component.

18. The method of claim 1, comprising computing a fast fourier transform of said detected output waveform.

19. The method of claim 1, further comprising computing a joint time-frequency transform of said detected output waveform.

20. The method of claim 1, wherein said input waveform comprises a plurality of components, each having a different frequency.

21. The method of claim 1, further comprising fitting said harmonic component to a first curve and a second curve, wherein said first curve describes a Faradaic signal and said second curve describes a background signal.

22. The method of claim 21, wherein said first curve is based, at least in part, on a modified Gaussian distribution.

23. The method of claim 21, wherein said second curve is a fifth order polynomial.

24. The method of claim 21, wherein said fitting comprises minimizing a mean square error.

25. The method of claim 23, wherein said fitting comprises using singular value decomposition.

26. The method of claim 1, wherein said analyzing comprises digital filtering.

27. The method of claim 26, wherein said filtering utilizes a filter chosen from the group consisting of a match filter, a weiner filter, and a Kalman filter.