초록 ▼

Improved hybridization probe compositions and method for detecting nucleic acids of interest are provided. A hybridization probe constructed to contain thionucleotides and a detectable label is hybridized to a nucleotide sequence of interest in the absence of nonpolar solvents, reducing agents and v

Improved hybridization probe compositions and method for detecting nucleic acids of interest are provided. A hybridization probe constructed to contain thionucleotides and a detectable label is hybridized to a nucleotide sequence of interest in the absence of nonpolar solvents, reducing agents and volume exclusion agents. The thionucleotides serve to increase the mass of bound probe molecules, thereby amplifying the signal available to detect the presence of the nucleic acid sequence of interest. Certain cations interact specifically with the thiol moiety of phosphorothioate groups of nucelic acid molecules in a manner that can be exploited to amplify probe binding or convey a detectable label. Thus, the mass of bound probe molecules can be further increased by the presence of specified multivalent cations from the transition metal group, such as Cu2+or Fe3+, during the annealing step.

대표청구항 ▼

A method for detecting the presence of specific nucleic acid base sequences in a sample suspected of containing said sequences in single-stranded form comprising: providing nucleic acid probe molecules which in single-stranded form are capable of binding specifically to said suspected nucleic acid b

A method for detecting the presence of specific nucleic acid base sequences in a sample suspected of containing said sequences in single-stranded form comprising: providing nucleic acid probe molecules which in single-stranded form are capable of binding specifically to said suspected nucleic acid base sequences and contain at least one thionucleotide; converting any double-stranded probe molecules to single-stranded form; annealing said single-stranded, thionucleotide-containing probe molecules to said suspected nucleic acid base sequences in the presence of a concentration of multivalent cations capable of promoting the formation of ionic interactions between phosphorothioate groups of said probe molecules; subsequent to said annealing, detecting the presence of any said probe molecules annealed to said suspected nucleic acid base sequences; and prior to said detecting, providing said probe molecules with a detectable label to provide a means for later detecting the presence of any said probe molecules annealed to said suspected nucleic acid base sequences.