METHOD AND ARRANGEMENT FOR CALIBRATING A SENSOR ELEMENT

• US 20100248243A1
• Filed: 10/29/2008
• Published: 09/30/2010
• Est. Priority Date: 11/20/2007
• Status: Active Grant

• Current Assignee: Boehringer Ingelheim Vetmedica GmbH (C.H. Boehringer Sohn AG & Co. KG)
• Original Assignee: Boehringer Ingelheim Vetmedica GmbH (C.H. Boehringer Sohn AG & Co. KG)
• Inventors: Friedrich, Katja, Weber, Renee, Paulicka, Peter, Gumbrecht, Walter, Stanzel, Manfred
• Granted Patent: US 10,081,831 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 2527/107 Temperature of melting, i.e. Tm

Abstract

A method is disclosed for calibrating a sensor element, which has an immobilized probe oligonucleotide, via which the bonding of a target nucleic acid (Z) can be detected by the sensor. In at least one embodiment, the method includes: a) bringing the sensor element into contact with a control nucleic acid (K), the melting temperature Tm(K) of which is less than the melting temperature Tm(Z) of the target nucleic acid (Z); b) hybridizing the control nucleic acid (K) to the probe oligonucleotide at a temperature T[p]<Tm(K), and detecting a positive control signal; and optionally c) modifying the stringent conditions such that T[n]>Tm(K) and detecting a negative control signal at a temperature T[n]. According to a refinement of at least one embodiment of the invention, a measuring signal of the target nucleic acid (Z) is measured at a measuring temperature T [mess], where Tm(K)<T[mess]<Tm(Z). The method is suited in particular for the calibration and quality control of microarrays.

Claims

1. A method for a sensor element including an immobilized probe oligonucleotide, binding of a target nucleic acid-(Z) being recordable via the sensor element, comprising:

a) contacting the sensor element with a control nucleic acid (K), the melting temperature of which, Tm(K), is lower than the melting temperature of the target nucleic acid, Tm(Z); and

b) hybridizing the control nucleic acid (K) to the probe oligonucleotide at a temperature T[p]<

Tm(K), and recording a positive control signal.

2. The method as claimed in claim 1, further comprising:

c) modifying the stringency conditions such that T[n] >

Tm(K), and recording a negative control signal.

3. The method claimed in claim 1, wherein

a) the contacting includes contacting the sensor element with a mixture including control nucleic acid (K) and target nucleic acid (Z), melting temperature of the control nucleic acid, Tm(K), being lower than the melting temperature of the target nucleic acid, Tm(Z);

and

b) the hybridizing includes,

b1) hybridizing the mixture to the probe oligonucleotide at a temperature T[p]<

Tm(K), and recording a positive control signal; and

b2) modifying the stringency conditions at a measuring temperature T=T[mess] such that;

Tm(K)<

T[mess]<

Tm(Z),

and recording a measurement signal.

4. The method as claimed in claim 2, further comprising:

c) modifying the stringency conditions such that T[n]>

Tm(Z), and recording a negative control signal.

5. The method as claimed in claim 1, wherein the stringency conditions are modified by increasing the temperature.

6. The method as claimed in claim 3, wherein a plurality of measurement signals, 1 to n, are measured at different temperatures T[mess 1−

n], where Tm(K)<

1−

n]<

Tm(Z).

7. The method as claimed in claims 1, wherein the method is carried out for a majority of sensor elements arranged in an array arrangement.

8. The method as claimed in claim 1, wherein target nucleic acid and control nucleic acid are labeled with a detectable label.

9. The method as claimed in claim 8, wherein the detectable label is an enzymatic label capable of converting a substrate into a product detectable by the sensor element.

10. The method as claimed in claim 1, wherein step (a) is preceded by contacting the sensor element with detectable product in order to record a first calibration signal.

11. The method as claimed in claim 1, wherein additionally a temperature compensation signal is recorded at each of the temperatures T[p], T[mess], T[mess 1−

n] and T[n] on a further sensor element on which detectable label is immobilized directly.

12. The method as claimed in claim 1, wherein the sensor element is an electrochemical sensor element, and wherein target nucleic acid and control nucleic acid are labeled with a detectable label which is an enzymatic label converting a substrate into a product detectable by the sensor element.

13. The method as claimed in claim 1, wherein the melting temperature Tm(K) of the control nucleic acid (K) is at least 5°

C. lower than the melting temperature Tm(Z) of the target nucleic acid (Z).

26. The method as claimed in claim 2, wherein the method is carried out for a majority of sensor elements arranged in an array arrangement.

27. The method as claimed in claim 2, wherein target nucleic acid and control nucleic acid are labeled with a detectable label.

14. An arrangement, comprising

at least one sensor element including a probe nucleotide immobilized thereon, binding of a target nucleic acid (Z) being recordable via the at least one sensor element, wherein a binding complex of probe oligonucleotide and target nucleic acid has a melting temperature Tm(Z);

at least one of heating and cooling means for changing the temperature at the at least one sensor element; and

at least one control nucleic acid capable of forming with the probe nucleotide a binding complex which has a melting temperature of Tm(K)<

Tm(Z).

15. The arrangement as claimed in claim 14, further comprising a majority of sensor elements arranged in an array arrangement.

17. The arrangement as claimed in either of claims 14, wherein the arrangement is a kit comprising an apparatus which comprises the at least one of heating and/or cooling means and the at least one sensor element, and further comprising a composition having the control nucleic acid.

18. The arrangement as claimed in claims 14, wherein the control nucleic acid is labeled with a detectable label.

19. The arrangement as claimed in claim 18, wherein the detectable label is an enzymatic label capable of converting a substrate into a product detectable by the sensor element.

20. The arrangement as claimed in claims 14, wherein detectable label is immobilized directly on a further sensor element.

21. The arrangement as claimed in claims 14, wherein the at least one sensor element is an electrochemical sensor element, and wherein the control nucleic acid is labeled with a detectable label which is an enzymatic label converting a substrate into a product detectable by the sensor element.

22. The arrangement as claimed in claims 14, wherein the melting temperature Tm(K) of the control nucleic acid (K) is at least 5°

C. lower than the melting temperature Tm(Z) of the target nucleic acid (Z).

23. The arrangement as claimed in claims 14, further comprising as control nucleic acid, a nucleic acid composition having randomized deca-oligonucleotide sequences.

24. The arrangement as claimed in claim 23, wherein the nucleic acids are biotinylated with randomized deca-oligonucleotide sequences.

25. The arrangement as claimed in claims 14, wherein the sensor element is an electrochemical sensor element.

28. The arrangement as claimed in claim 15, wherein the control nucleic acid is stored by way of a dry reagent in the arrangement.

29. The arrangement as claimed in claim 15, wherein the arrangement is a kit comprising an apparatus which comprises the at least one of heating and cooling means and the at least one sensor element, and further comprising a composition having the control nucleic acid.

16. The arrangement as claimed in, wherein the control nucleic acid is stored by way of a dry reagent in the arrangement.