Assay for genetic polymorphisms using scattered light detectable labels
AssigneeInvitrogen Corporation
InventorsGary Bee, David E. Kohne, Linda Korb, Todd Peterson, Juan Yguerabide
Life of this patent
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Abstract
Described are methods for determining the presence or absence of particular polymorphisms in CYP2D6 and other genes using scattered light detectable particles as detectable labels, and compositions useful in such methods.
Claims27
A method for determining the presence or absence of a CYP2D6 target sequence in a sample of DNA containing nucleic acid corresponding to CYP2D6, comprising
- contacting said nucleic acid with a probe under stringent binding conditions, and detecting the presence or absence of target sequence bound with said probe, wherein said target sequence or said probe is bound with scattered light detectable particle, and said detecting comprises observing light scattered from said particle as an indication of said presence or absence.
-15. (canceled)
An allele specific probe, comprising a molecule that preferentially binds to a labeled CYP2D6 target nucleic acid sequence at least partially comprising a sequence corresponding to a polymorphism in a CYP2D6 gene, wherein a scattered light detectable particle 1-500 nm in size is bound with said probe.
The probe of claim 20, wherein said probe comprises a nucleotide sequence that is designed to discriminate different allelic forms of at least one CYP2D6 target nucleic acid sequence.
-26. (canceled)
An isolated CYP2D6 nucleic acid sequence, wherein said nucleic acid sequence is bound with a probe and said nucleic acid sequence or said probe is bound with a scattered light detectable particle.
-58. (canceled)
A method for detecting specifically an allele of a pharmacogenetically relevant gene involved in drug metabolism in a sample, said allele comprising a target nucleotide sequence that is unique to said allele, said method comprising the steps of:
- (a) contacting said allele of said pharmacogenetically relevant gene involved in drug metabolism in said sample with a nucleic acid probe under differential hybridization conditions that allow said nucleic acid probe to hybridize specifically to a nucleic acid molecule in said allele of said pharmacogenetically relevant gene involved in drug metabolism in said sample, wherein said nucleic acid molecule comprises said target nucleotide sequence, and wherein either said nucleic acid probe or said nucleic acid molecule is labeled with one or more scattered-light detectable particles of a size between 1 and 500 nm inclusive, thereby forming hybridized nucleic acid molecules that are labeled;
- (b) illuminating said one or more scattered-light detectable particles bound to said hybridized nucleic acid molecules under conditions which produce scattered light from said one or more scattered-light detectable particles and in which light scattered from said one or more scattered-light detectable particles can be detected by a human eye with less than 500 times magnification and without electronic amplification;
- (c) detecting light scattered by said one or more scattered-light detectable particles under said conditions as indicative of the presence of said allele of said pharmacogenetically relevant gene involved in drug metabolism in said sample;
- and
- (d) contacting said allele of said pharmacogenetically relevant gene involved in drug metabolism in said sample with a capture probe (i) that is immobilized on a solid surface and (ii) that hybridizes to said nucleic acid molecule comprising said target nucleotide sequence, wherein said nucleic acid probe is labeled with scattered-light detectable particles.
Dependencies are inferred from the claim text.