| abstract |
The invention relates to methods and their implementation material based on the discovery that RNA replicase, such as QB replicase, exhibits DNA-dependent RNA polymerase ("DDRP") activity with d segments nucleic acid comprising chimeric segments of DNA and DNA: RNA, which comprise a 2'-deoxyribonucleotide and present RNA sequences replicable autocatalytically by replicase. The invention relates to the use of amplification methods in the detection of nucleic acid analytes, such as in hybridization assays of nucleic acid probes in which a nucleic acid analyte is hybridized with one or more nucleic acid probes, which are processed to generate a DNA segment capable of being amplified by the DDRP activity of an RNA replicase of an RNA replicated autocatalytically in order to produce a large number of easily detectable reporter molecules. The method allows the replacement of an RNA in the presence of MN + 2, Co + 2 or Zn + 2 in the solution in which the DDRP activity takes place. |