Monitoring oligonucleotide binding processes using chemiluminescence quenching
First Claim
1. A method for detecting and/or quantifying a target nucleic acid sequence in a sample comprising:
- contacting said sample with an oligonucleotide sequence containing at least a probe sequence capable of binding to said target nucleic acid sequence, wherein said oligonucleotide sequence is labelled with each of at least one chemiluminescent molecule and at least one quencher molecule capable of attenuating chemiluminescence from said chemiluminescent molecule, the chemiluminescent and quencher molecules being arranged so that the interaction thereof changes according to whether the probe sequence is bound to said target nucleic acid sequence, such that in one of the bound and unbound states said chemiluminescence is substantially attenuated, and in the other thereof there is reduced or no attenuation of the chemiluminescence;
chemically activating the chemiluminescent molecule; and
detecting the chemiluminescence emission from said chemiluminescent molecule and comparing said chemiluminescence emission with that corresponding to the absence of said target nucleic acid sequence thereby detecting and/or quantifying said target nucleic acid sequence.
7 Assignments
0 Petitions
Accused Products
Abstract
Oligonucleotide building processes are monitored by means of an oligonucleotide probe which in one embodiment is labelled at one end with a chemiluminescent label and at the other end with a quencher molecule. The conformation of the oligonucleotide probe changes according to whether the probe hybridises with a substantially complementary nucleic acid sequence. In the non-hybridised state the chemiluminescent label is sufficiently close proximity to the quencher that the chemiluminescent emission is substantially attenuated, but in the hybridised state the separation is such that there is little or no attenuation. Particular probes and emitter/quencher pairs are disclosed.
22 Citations
26 Claims
-
1. A method for detecting and/or quantifying a target nucleic acid sequence in a sample comprising:
-
contacting said sample with an oligonucleotide sequence containing at least a probe sequence capable of binding to said target nucleic acid sequence, wherein said oligonucleotide sequence is labelled with each of at least one chemiluminescent molecule and at least one quencher molecule capable of attenuating chemiluminescence from said chemiluminescent molecule, the chemiluminescent and quencher molecules being arranged so that the interaction thereof changes according to whether the probe sequence is bound to said target nucleic acid sequence, such that in one of the bound and unbound states said chemiluminescence is substantially attenuated, and in the other thereof there is reduced or no attenuation of the chemiluminescence; chemically activating the chemiluminescent molecule; and detecting the chemiluminescence emission from said chemiluminescent molecule and comparing said chemiluminescence emission with that corresponding to the absence of said target nucleic acid sequence thereby detecting and/or quantifying said target nucleic acid sequence. - View Dependent Claims (2, 3, 4, 5, 6, 7, 10, 13, 14, 15, 16, 17, 18, 19)
-
-
8. A method for detecting and/or quantifying a target nucleic acid sequence in a sample comprising:
-
contacting said sample with at least two oligonucleotides having different sequences capable of simultaneously binding to said target nucleic acid sequence, wherein said at least two oligonucleotides are labelled respectively with at least one chemiluminescent molecule and at least one quencher molecule capable of attenuating chemiluminescence when said two labelled oligonucleotides bind to said target nucleic acid sequence substantially adjacent to each other; chemically activating the chemiluminescent molecule; and detecting the chemiluminescence emission from said chemiluminescent molecule and comparing said chemiluminescence emission with that corresponding to the absence of said target nucleic acid sequence thereby detecting and/or quantifying said target nucleic acid sequence. - View Dependent Claims (11, 12)
-
-
9. A method for detecting and/or quantifying a target nucleic acid sequence in a sample comprising:
-
contacting said sample with at least two oligonucleotides capable of binding to each other, wherein said at least two oligonucleotides are labelled respectively with at least one chemiluminescent molecule and at least one quencher molecule capable of attenuating chemiluminescence when said labelled oligonucleotides are bound to each other, and wherein there is little or no attenuation of said chemiluminescence from the chemiluminescent molecule when the binding of said labelled oligonucleotides to each other is inhibited by the presence of said target nucleic acid sequence; chemically activating the chemiluminescent molecule; and detecting the chemiluminescence emission from said chemiluminescent molecule and comparing said chemiluminescence emission obtained in the presence and absence of said target nucleic acid sequence thereby detecting and/or quantifying said target nucleic acid sequence.
-
- 20. A labelled oligonucleotide for detecting and/or quantifying a target nucleic acid sequence comprising an oligonucleotide probe sequence capable of binding to said target nucleic acid sequence, said oligonucleotide being labelled with a chemiluminescent molecule requiring chemical activation and a quencher molecule and being arranged such that the quencher molecule and the chemiluminescent molecule requiring chemical activation interact to cause a change in a subsequent chemiluminescent emission according to whether the labelled oligonucleotide is bound to said target nucleic acid sequence.
Specification