Compositions and methods for the simultaneous detection and quantification of multiple specific nucleic acid sequences
AssigneeGen-Probe Incorporated
InventorsNorman Charles Nelson, James Stuart Woodhead, Ian Weeks, Azzouz Ben Cheikh
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Abstract
The invention relates to methods for simultaneously or sequentially detecting multiple nucleic acid analytes in a single medium utilizing oligonucleotide hybridization probes coupled to different chemiluminescent labeling reagents. The methods may be used in a heterogeneous, homogeneous or non-homogeneous assay system. The invention also relates to specific combinations of chemiluminescent labeling reagents suitable, when coupled to an oligonucleotide probe, for use together in methods for the detection of multiple nucleic acid analytes. The invention also concerns kits useful in these methods.
Claims24
A kit for the detection of two or more nucleic acid analytes suspected of being present in a sample, comprising, in one or more container:
- a) at least two different oligonucleotide hybridization assay probes, each able to specifically hybridize to a different nucleic acid target analyte,
- b) at least one different chemiluminescent label coupled to each said probe, wherein each chemiluminescent label in said kit is chosen so that the chemiluminescence emitted by said label upon induction of light emission is separately distinguishable on the basis of wavelength of light emission, kinetics of the chemiluminescent reaction, or pH of reaction from the chemiluminescence emitted by at least one other of said labels;
- and said coupled labels are able to react with a destabilizing agent which will selectively destroy or inhibit the chemiluminescent potential of said labels when said probe has not formed a stable nucleic acid hybrid with an analyte without extinguishing the chemiluminescent potential of said labels coupled to hybridized probe;
- and the rate of destruction or inhibition of at least one hybrid-unassociated coupled label by said destabilizing agent is no more than 50 fold the rate of destruction or inhibition of at least one other of said hybrid-unassociated coupled labels by said agent.
Dependencies are inferred from the claim text.