| abstract |
The present invention entails methods, and kits for carrying them out, based on the discovery that an RNA replicase, such as QB replicase, has DNA-dependent RNA polymerase ('DDRP') activity with nucleic acid segments including DNA and DNA:RNA chimeric segments, which comprise a 2'-deoxyribonucleotide and which have sequences of RNAs that are autocatalytically replicable by the replicase. The invention entails use of the amplification methods in detecting nucleic acid analytes, as in nucleic acid probe hybridization assays wherein a nucleic acid analyte is hybridized with one or more nucleic acid probes, which are processed to generate a DNA segment which is amplifiable through the DDRP activity of an RNA replicase of an autocatalytically replicated to provide an abundance of readily detectable reporter molecules. The method permits the replacement of an RNA in presence of Mn?+2, Co+2, or Zn+2¿ in the solution in which the DDRP activity occurs. |