Device and method of detecting mutations and polymorphisms in DNA
First Claim
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1. A method of detecting a mutation in a target DNA comprising:
- obtaining a first measurement of a resonance parameter of a resonator sensor, said resonator sensor having a vibrating element, said vibrating element having a surface, and a probe DNA immobilized on said surface, said probe DNA hybridized with a target DNA;
contacting the hybridized probe DNA and target DNA with a mismatch binding molecule in order to form a mismatch binding molecule/duplex DNA complex;
obtaining a second measurement of the resonance parameter of the resonator sensor;
comparing the first and second measurement of the resonance parameter to detect formation of the mismatch binding molecule/duplex DNA complex;
determining if the mismatch binding molecule bound to the duplex DNA due to binding to a mismatch pairing or if the mismatch binding molecule bound to the duplex DNA due to nonspecific interactions with the duplex DNA backbone; and
detecting the presence of a mutation in the target DNA when it is determined that the mismatch binding molecule bound to the duplex DNA due to binding to a mismatch pairing.
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Abstract
There is provided a resonator sensor useful for detecting polymorphisms and mutations in DNA. The resonator sensor has a capture molecule immobilised on its surface, the capture molecule being either a probe DNA containing a reference sequence, or a mismatch binding molecule, and being capable of forming a probe DNA/target DNA/mismatch binding molecule complex on the surface of the resonator. A method for detecting mutations in a target DNA, including single nucleotide polymorphisms, is also provided.
78 Citations
42 Claims
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1. A method of detecting a mutation in a target DNA comprising:
- obtaining a first measurement of a resonance parameter of a resonator sensor, said resonator sensor having a vibrating element, said vibrating element having a surface, and a probe DNA immobilized on said surface, said probe DNA hybridized with a target DNA;
contacting the hybridized probe DNA and target DNA with a mismatch binding molecule in order to form a mismatch binding molecule/duplex DNA complex;
obtaining a second measurement of the resonance parameter of the resonator sensor;
comparing the first and second measurement of the resonance parameter to detect formation of the mismatch binding molecule/duplex DNA complex;
determining if the mismatch binding molecule bound to the duplex DNA due to binding to a mismatch pairing or if the mismatch binding molecule bound to the duplex DNA due to nonspecific interactions with the duplex DNA backbone; and
detecting the presence of a mutation in the target DNA when it is determined that the mismatch binding molecule bound to the duplex DNA due to binding to a mismatch pairing. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
- obtaining a first measurement of a resonance parameter of a resonator sensor, said resonator sensor having a vibrating element, said vibrating element having a surface, and a probe DNA immobilized on said surface, said probe DNA hybridized with a target DNA;
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21. A method of detecting a mutation in a target DNA comprising:
- obtaining a first measurement of a resonance parameter of a resonator sensor, said resonator sensor having a vibrating element, said vibrating element having a surface, and a mismatch binding molecule immobilized on said surface;
contacting the mismatch binding molecule with a hybridized probe DNA and target DNA in order to form a mismatch binding molecule/duplex DNA complex;
obtaining a second measurement of the resonance parameter of the resonator sensor;
comparing the first and second measurement of the resonance parameter to detect formation of the mismatch binding molecule/duplex DNA complex;
determining if the mismatch binding molecule bound to the duplex DNA due to binding to a mismatch pairing or if the mismatch binding molecule bound to the duplex DNA due to nonspecific interactions with the duplex DNA backbone; and
detecting the presence of a mutation in the target DNA when it is determined that the mismatch binding molecule bound to the duplex DNA due to binding to a mismatch pairing. - View Dependent Claims (22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42)
- obtaining a first measurement of a resonance parameter of a resonator sensor, said resonator sensor having a vibrating element, said vibrating element having a surface, and a mismatch binding molecule immobilized on said surface;
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