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 wherein the said oligonucleotide 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, 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; and
changing the conditions so as to cause the chemiluminescent molecule to undergo a chemiluminescent reaction; and
monitoring the chemiluminescence emission and comparing said emission with that corresponding to the absence of said target nucleic acid.
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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.
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Citations
26 Claims
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1. A method for detecting and/or quantifying a target nucleic acid sequence in a sample comprising:
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contacting said sample with an oligonucleotide sequence containing at least a probe sequence capable of binding to said target nucleic acid wherein the said oligonucleotide 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, 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; and
changing the conditions so as to cause the chemiluminescent molecule to undergo a chemiluminescent reaction; and
monitoring the chemiluminescence emission and comparing said emission with that corresponding to the absence of said target nucleic acid. - View Dependent Claims (2, 3, 4, 5, 6, 7, 10, 13, 14, 15, 16, 17, 18, 19)
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8. A method for detecting and/or quantifying a target nucleic acid in a sample comprising:
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contacting said sample with at least two oligonucleotides having different sequences capable of simultaneous binding to said target nucleic acid and where said at least two oligonucleotides are each labelled respectively with at least one chemiluminescent emitting molecule and at least one quencher molecule capable of attenuating chemiluminescence when said two labelled oligonucleotides bind to the said target nucleic acid substantially adjacent to each other; and
changing the conditions so as to cause the chemiluminescent molecule to undergo a chemiluminescent reaction; and
monitoring the chemiluminescence emission and comparing said emission with that corresponding to the absence of said target nucleic acid. - View Dependent Claims (11, 12)
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9. A method for detecting and/or quantifying a target nucleic acid in a sample comprising:
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contacting said sample with at least two oligonucleotides capable of binding to each other and where at least two of said oligonucleotides are each labelled respectively with at least one chemiluminescent emitting 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; and
changing the conditions so as to cause the chemiluminescent molecule to undergo a chemiluminescent reaction; and
monitoring the chemiluminescence emission and comparing the said emission obtained in the presence and absence respectively of said target nucleic acid.
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- 20. A labelled oligonucleotide for detecting and/or quantifying a target nucleic acid, comprising an oligonucleotide probe capable of binding to said target nucleic acid, said oligonucleotide being labelled with a chemiluminescent molecule and a quencher molecule and being arranged such that the said quencher molecule and the chemiluminescent molecule 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