| abstract |
The present invention provides a method for detecting the presence of a target polynucleotide in a sample. The method comprises the steps of (a) contacting the sample under hybridizing conditions with (i) a single-stranded polynucleotide probe capable of hybridizing to the target polynucleotide and comprising a polynucleotide and at least one intercalating molecule attached to a nucleotide of the polynucleotide by means of a linker arm, and (ii) a background-reducing reagent which chemically modifies the intercalating molecule when the probe to which it is attached is single-stranded; and (b) detecting a property change resulting from the intercalation of the intercalating molecule into a target-probe hybrid, thereby detecting the target polynucleotide. The intercalating molecule which is part of the polynucleotide probe induces a change in a property, in either the probe, the target polynucleotide or a target-probe hybrid. The property change can be detected, for example, by means of a generated signal which can be identified or quantified. The present invention can be employed in a heterogeneous (two step or two phase) assay using a support to immobilize the target or probe, and a washing step, and in a homogeneous (one step or one phase) assay using a hybridization solution. Also provided are a composition and a nucleic acid hybridization kit useful for detecting the presence of a target polynucleotide in a sample. |