Multiple fluorophore detector system
AssigneeBecton, Dickinson and Company
InventorMatthew Collis
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Abstract
A detector oligonucleotide comprises multiple pairs of a donor fluorophore and a quencher molecule, which donor fluorophores and quencher molecules are separated by a site that is capable of being cleaved when in double-stranded form. The detector oligonucleotide may be made double-stranded in a manner that depends on the presence of a target nucleic acid, allowing the cleavage sites to be cleaved. Separation of the donor fluorophores and the quencher molecules decreases fluorescence quenching and generates a detectable change in a fluorescence parameter of the fluorophores of the detector oligonucleotide. By using multiple donor/quencher pairs, the present detector oligonucleotide advantageously generates a high signal to noise ratio and high efficiency in detection of a target nucleic acid.
Claims20
A detector oligonucleotide, comprising at least two pairs of a donor fluorophore and quencher molecules, the donor fluorophore and quencher molecule being in close proximity such that otherwise detectable signal from said donor fluorophore is quenched by said quencher molecule, wherein said donor fluorophore and said quencher molecule in each said pair are separated by a cleavage site that is cleavable when hybridized to a target in a double-stranded form, and wherein cleavage at said cleavage site is capable of creating a detectable signal by the donor fluorophore that indicates the presence of a target nucleic acid.
A single-stranded first detector oligonucleotide and a single-stranded second oligonucleotide, wherein the first detector oligonucleotide comprises a plurality of pairs donor fluorophore and quencher molecules, wherein the donor fluorophore and quencher molecule in each said pair are separated by a cleavage site that is cleavable when hybridized to a target in a double-stranded form, and wherein the cleavage site is double-stranded when the first detector oligonucleotide forms a duplex with a second oligonucleotide that formed in the presence of a target nucleic acid.
A partially double-stranded detector oligonucleotide, comprising a plurality of pairs of donor fluorophores and quencher molecule, wherein the donor fluorophore and quencher molecule in each said pair is in close proximity, wherein said donor fluorophore and said quencher molecule in each said pair is separated by a cleavage side, wherein said partially double-stranded detector oligonucleotide comprises a single-stranded portion that is capable of forming a duplex with a target nucleic acid.
A kit, comprising a single-stranded first detector oligonucleotide, comprising a plurality of pairs of a donor fluorophores and a quencher molecules, wherein the donor fluorophore and quencher molecule are in close proximity, wherein said donor fluorophore and said quencher molecule in each said pair are separated by a cleavage site that is cleavable when hybridized to a target in a double-stranded form, and wherein the cleavage site is double-stranded when the first detector oligonucleotide forms a duplex with a second oligonucleotide capable of being formed in the presence of a target nucleic acid.
Dependencies are inferred from the claim text.