Bridged element for detection of a target substance

• US 8,078,408 B2
• Filed: 08/06/2004
• Issued: 12/13/2011
• Est. Priority Date: 08/06/2003
• Status: Active Grant

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: The Procter & Gamble Company
• Inventors: Albert, Fred, Wright, Brad
• Primary Examiner(s): Lin, Jerry
• Application Number:
  US10/913,021
• Publication Number:
  US 20050074871A1
• Time in Patent Office: 2,685 Days
• Field of Search: 702/19
• US Class Current: 702/19
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2523/307 Denaturation or renaturation by electric current/voltage
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings

Abstract

Physical changes resulting from an association between a template molecule and a target molecule are detected by monitoring changes in the template molecule. Exemplary changes include a change in a physical dimension or stiffness of the template molecule, a change in electrical conductivity of the template molecule and a change in the energy required to dissociate the target molecule and the template molecule. The magnitude of the change is indicative of the specific identity of the target molecule.

Claims

1. A device comprising:

a template molecule linked between at least two contact points including a first contact point disposed on a first surface and a second contact point disposed on a second surface, wherein the first surface is independent of the second surface, and wherein the template molecule includes at least one recognition site for a target molecule; and

a detector coupled to the first contact point and the second contact point, the first contact point and the second contact point having a first physical parameter, the detector configured to generate an output signal based on a change in the first physical parameter indicative of electrical conduction through the template molecule, wherein the change in the first physical parameter corresponds to an association between the target molecule and the template molecule, wherein a single stranded conductive nucleic acid molecule is selected as the template molecule; and

wherein the template molecule, the first contact point, the detector and the second contact point form an electrical circuit, and further, wherein the detector comprises a driving circuit for providing a current or voltage through the electrical circuit for disassociating the template molecule from the target molecule.

2. The device of claim 1 wherein the template molecule is ssDNA or ssRNA.

3. The device of claim 1 wherein the target molecule is a nucleic acid comprising a sequence sufficiently complementary to a sequence in the template molecule so that the target molecule will hybridize to said template molecule.

4. The device of claim 1 wherein the template molecule is a nucleic acid that is bonded to the first contact point at its 3′

-end and to the second contact point at its 5′

- end.

5. The device of claim 1 further comprising a second physical parameter which includes at least one of:

a resonant frequency of the first contact point relative to a reference point;

a resonant amplitude of the first contact point relative to the reference point;

a distance between the first contact point and the reference point; and

an alignment between the first contact point and the reference point.

6. The device of claim 5 wherein the reference point includes the second contact point.

7. The device of claim 1 wherein the first surface includes a movable surface.

8. The device of claim 1 wherein the detector includes at least one of an optical sensor, a magnetic field sensor, an electric field sensor, a capacitance sensor, a resistance sensor and a strain sensor.

9. The device of claim 1 wherein the detector includes at least one of a comparator and a bridge circuit.

10. The device of claim 1 wherein the detector includes a resonance driver in communication with the first surface.

11. The device of claim 5 wherein the detector is configured to generate the output signal based on a change in the second physical parameter.

12. The device of claim 1 wherein the detector is coupled to the second contact point and wherein the first contact point having a first physical parameter includes the first contact point and the second contact point having a first electrical parameter.

13. The device of claim 12 wherein the detector includes a voltage source coupled to the first contact point and the second contact point and wherein the first electrical parameter includes a measure of a current.

14. The device of claim 13 wherein the voltage source is configured to supply an increasing potential.

15. The device of claim 12 wherein the detector includes a current source coupled to the first contact point and the second contact point and wherein the first electrical parameter includes a measure of a voltage.

16. The device of claim 15 wherein the current source is configured to supply an increasing current.

17. The device of claim 12 wherein the detector includes a driver circuit configured to deliver an electrical signal and wherein the change in the first physical parameter corresponds to a disbonding of the target molecule and the template molecule.

18. The device of claim 1 wherein at least one of the first surface and the second surface includes at least one of glass, quartz, silicon and a polymer.

19. The device of claim 1 wherein the template molecule is attached to the contact point with at least one of thiol, gold, biotin or streptavidin.

31. The device of claim 1 wherein an increase in the first physical parameter corresponds to an association between the target nucleic acid molecule and the template nucleic acid molecule.

36. The device of claim 1 wherein the output signal correlates to the percent nucleic acid sequence identity between the template molecule and the target molecule.

20. A method comprising:

exposing a target molecule to a template molecule to form a hybridized nucleic acid molecule, the template molecule linked between at least two contact points including a first contact point disposed on a first surface and a second contact point disposed on a second surface, wherein the template molecule includes at least one recognition site for a target molecule, wherein a single stranded conductive nucleic acid molecule is selected as the template molecule;

generating an output signal as a function of a change in a first physical parameter indicative of electrical conduction through the template molecule and measured using the first contact point relative to a reference point, wherein the change in the first physical parameter corresponds to an association between the target molecule and the template molecule; and

generating a current or voltage that passes through the first contact point, the template molecule and the second contact point to disassociate the hybridized nucleic acid molecule.

21. The method of claim 20 further including:

monitoring a resonant frequency of the first contact point relative to a reference point;

monitoring a resonant amplitude of the first contact point relative to the reference point;

monitoring a distance between the first contact point and the reference point; and

monitoring an alignment between the first contact point and the reference point.

22. The method of claim 21 wherein the reference point includes the second contact point.

23. The method of claim 20 further including applying an electrical signal to the template molecule to disbond the target molecule.

37. The method of claim 20 wherein the output signal correlates to the percent nucleic acid sequence identity between the template molecule and the target molecule.

24. A system comprising:

a target molecule introduction port for receiving a sample;

a sensor having a template molecule in communication with the target molecule introduction port, the template molecule linked between a first point on a first surface and a second point on a second surface, the first surface independent of the second surface and wherein the template molecule has at least one recognition site specific to a target molecule, wherein a single stranded conductive nucleic acid molecule is selected as the template molecule;

a detector coupled to the first point and the second point and configured to generate an output signal based on a change in a measured parameter corresponding to an association between the template molecule with the target molecule, wherein the measured parameter is electrical conduction through the template molecule; and

an output circuit to provide a result based on the output signal; and

wherein the template molecule, the first contact point, the detector and the second contact point form an electrical circuit, and further, wherein the detector comprises a driving circuit for providing a current or voltage through the electrical circuit for disassociation of the template molecule from the target molecule.

25. The system of claim 24 further including a plurality of sensors, each sensor coupled to the detector by a multiplexer.

26. The system of claim 24 a processor coupled to the detector and having access to a memory, wherein the memory provides data storage for identifying a target molecule based on the change in the measured parameter.

27. The system of claim 24 wherein the output circuit includes at least one of an interface, a display and a wireless transceiver.

28. The system of claim 24 further including a test circuit coupled to the template molecule to determine conductivity of the template molecule.

29. The system of claim 24 further including a reset circuit coupled to the template molecule to disbond a target molecule from the template molecule.

30. The system of claim 24 further including a housing for containment of at least one of the target molecule introduction port, the sensor, the detector and the output circuit.

38. The system of claim 24 wherein the output signal correlates to the percent nucleic acid sequence identity between the template molecule and the target molecule.

32. A device comprising:

a template nucleic acid molecule linked between at least two contact points including a first contact point disposed on a first surface and a second contact point disposed on a second surface, wherein the first surface is independent of the second surface, and wherein the template nucleic acid molecule includes at least one recognition site for a target nucleic acid molecule and is selected as a conductive single stranded nucleic acid molecule;

wherein the template nucleic acid molecule, the first contact point, the detector and the second contact point form an electrical circuit, and further, wherein the detector comprises a driving circuit for providing a current or voltage through the electrical circuit for disassociation of the template molecule from the target molecule; and

a detector coupled to the first contact point and the second contact point, the first contact point and the second contact point having a first physical parameter indicative of electrical conduction though the template nucleic acid molecule, the detector configured to generate an output signal based on a change in electrical conduction through the template nucleic acid molecule associated with hybridization of a target nucleic acid molecule to the template nucleic acid molecule and with disassociation of the template nucleic acid molecule from the target nucleic acid molecule.

33. The device of claim 32 wherein the first and second surfaces are a fixed distance from each other.

34. A method comprising:

a) exposing a target nucleic acid molecule to a template nucleic acid molecule to yield a hybridized nucleic acid molecule, the template molecule linked between at least two contact points including a first contact point disposed on a first surface and a second contact point disposed on a second surface, wherein the template nucleic acid molecule includes at least one recognition site for the target nucleic acid molecule and is selected as a single stranded conductive nucleic acid molecule;

b) generating an output signal as a function of a change in a first physical parameter indicative of electrical conduction through the template molecule and measured using the first contact point relative to a reference point, wherein the change in the first physical parameter is associated with hybridization of the target nucleic acid molecule to the template nucleic acid molecule; and

c) generating a current or voltage that passes through the first contact point, the template molecule and the second contact point to disassociate the hybridized nucleic acid molecule.

35. The method of claim 34 wherein the change in the first physical parameter is correlated to the percent nucleic acid sequence identity between the target nucleic acid molecule and the template nucleic acid molecule.