Biosensor, device and method for detecting nucleic acids by means of at least two units for immobilizing nucleic acids

• US 20040152091A1
• Filed: 02/17/2004
• Published: 08/05/2004
• Est. Priority Date: 03/16/2001
• Status: Abandoned Application

• Current Assignee: Siemens AG, Teknologi & Produkt Udvikling A/S
• Original Assignee: Siemens AG
• Inventors: Schiente, Meinard, Thewes, Roland, Paulus, Christian
• Application Number:
  US10/472,074
• Publication Number:
  US 20040152091A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

A device and method for detecting nucleic acids. The device having a biosensor with at least two nucleic acid immobilization units and an electrical detection circuit. In the biosensor, the at least two nucleic acid immobilization units are in this case electrically conductive and electrically insulated from one another. The at least two nucleic acid immobilization units are provided with first nucleic acid molecules acting as scavenger molecules. The first nucleic acid molecules are present as single-stranded molecules and can bind second nucleic acid molecules to be detected. The first single-stranded nucleic acid molecules acting as scavenger molecules are provided with a redox-active label capable of generating a detectable signal. The electrical detection circuit is configured in such a way that it detects the hybridization even of the nucleic acid molecules with the scavenger molecules by means of the label.

Claims

1. A method for detecting nucleic acids by means of at least two nucleic acid immobilization units,

in which the at least two nucleic acid immobilization units are electrically conductive and electrically insulated from one another,

in which the at least two nucleic acid immobilization units are provided with first nucleic acid molecules acting as scavenger molecules, the first nucleic acid molecules being present as single-stranded molecules and being capable of binding second nucleic acid molecules to be detected, and the first single-stranded nucleic acid molecules acting as scavenger molecules being provided with a redox-active label which can generate a detectable signal,

in which a sample to be studied is brought in contact with the at least two nucleic acid immobilization units, wherein the sample to be studied may contain the second nucleic acid molecules to be detected,

in which second nucleic acid molecules contained in the sample to be studied become bound to the scavenger molecules, so that double-stranded hybrid molecules are formed,

in which the second nucleic acid molecules are detected by means of a signal caused by the redox-active label, by using an electrical detection circuit integrated in a semiconductor chip, and

in which an immobilization unit is selected for individual detection of the signal.

2. The method as claimed in claim 1, in which the signal generated by the redox-active label is detected by measuring a current flow, a resistance or a conductivity.

3. The method as claimed in claim 2, in which the redox-active label is a photo-inducible redox-active label or a chemically inducible redox-active label.

4. The method as claimed in claim 3, in which the photo-inducible redox-active label is a photosynthetic bacterial reaction center, a cyclophane or an at least bimolecular electron-donor/electron-acceptor complex.

5. The method as claimed in claim 4, in which the electron donor and the electron acceptor of the bimolecular electron-donor/electron-acceptor complex is a charge transfer complex or a transition metal complex.

6. The method as claimed in one of claims 1 to 5, in which DNA molecules, RNA molecules or PNA molecules are detected as the nucleic acids.

7. The method as claimed in claim 6,

in which DNA or RNA single strands with a predetermined nucleotide sequence are detected as the nucleic acid molecules, and

in which DNA probe molecules with a nucleotide sequence complementary to the predetermined nucleotide sequence are used as scavenger molecules.

8. The method as claimed in claim 7, in which unbound DNA probe molecules are removed from the at least two immobilization units.

9. The method as claimed in claim 8, in which an enzyme with nuclease activity is brought in contact with the immobilization unit in order to remove unbound DNA probe molecules.

10. The method as claimed in claim 9, in which at least one of the following substances is used as the enzyme with nuclease activity:

mung bean nuclease,

nuclease P1,

nuclease S1, or

DNA polymerases which are capable of degrading single-stranded DNA owing to their

5′

->

3′

exonuclease activity or their

3′

->

5′

exonuclease activity.

11. The method as claimed in one of claims 1 to 10, in which the immobilization units comprise gold.

12. The method as claimed in one of the preceding claims, in which the at least two immobilization units are arranged on a semiconductor chip.

13. The method as claimed in claim 12, in which the semiconductor chip is a CMOS chip.

14. A biosensor for detecting nucleic acids with at least two nucleic acid immobilization units and with an electrical detection circuit,

in which the at least two nucleic acid immobilization units are electrically conductive and electrically insulated from one another,

in which the at least two nucleic acid immobilization units are provided with first nucleic acid molecules acting as scavenger molecules, the first nucleic acid molecules being present as single-stranded molecules and being capable of binding second nucleic acid molecules to be detected, and the first single-stranded nucleic acid molecules acting as scavenger molecules being provided with a redox-active label which can generate a detectable signal, and

in which the electrical detection circuit is configured in such a way that the detection circuit detects nucleic acid molecules, which have bound to the scavenger molecules, by means of the label, and

in which the electrical detection circuit has selection electronics for individual selection of at least one immobilization unit, and in which the electrical detection circuit is integrated in a semiconductor chip.

15. The biosensor as claimed in claim 14, which has a plurality of nucleic acid immobilization units in a regular arrangement.

16. The biosensor as claimed in claim 15, in which the immobilization units are arranged on the semiconductor chip.

17. The biosensor as claimed in claim 16, in which the semiconductor chip is a CMOS chip.

18. The biosensor as claimed in one of claims 14 to 17, in which the electrical detection circuit has a preamplifier for preamplifying the detected signal for each immobilization unit.

19. The biosensor as claimed in one of claims 14 to 18, in which the electrical detection circuit has an analog/digital converter for converting the detected signal for each immobilization unit.

20. The biosensor as claimed in one of claims 14 to 19, in which the electrical detection circuit has an evaluation unit for the detected signal for each immobilization unit.

21. The biosensor as claimed in one of claims 14 to 20, in which the electrical evaluation unit for each immobilization unit has a unit for adding up the charge quantity imparted to the respective immobilization unit.

22. A device for detecting nucleic acids with a biosensor, which has at least two nucleic acid immobilization units and an electrical detection circuit, in the biosensor:

the at least two nucleic acid immobilization units being electrically conductive and electrically insulated from one another,

the at least two nucleic acid immobilization units being provided with first nucleic acid molecules acting as scavenger molecules, the first nucleic acid molecules being present as single-stranded molecules and being capable of binding second nucleic acid molecules to be detected, and the first single-stranded nucleic acid molecules acting as scavenger molecules being provided with a redox-active label which can generate an electrochemically detectable signal, and

the electrical detection circuit being configured in such a way that the detection circuit detects nucleic acid molecules, which have bound to the scavenger molecules, by means of the label, and

in which the electrical detection circuit has selection electronics for individual selection of at least one immobilization unit, and in which the electrical detection circuit is integrated in a semiconductor chip.