Diagnostic reagent, kit and method employing polynucleotide displacement, separation, enzymatic cleavage and adenosine phosphate detection

摘要

A reagent complex is disclosed containing (1) a probe polynucleotide having a target binding region complementary to a target nucleotide sequence and (2) a signal strand polynucleotide bound by base pairing to a portion of the target binding region. The signal strand is displaced from the reagent complex by the target nucleotide sequence and separated from intact reagent complexes. A digestable ribonucleotide segment of the displaced signal strand, such as a 3'+0, terminal segment, is digested to ribonucleotide phosphates, which are phosphorylated to ATP. The ribonucleotide phosphates or ATP are detected.