Nucleic acid amplification with DNA-dependent RNA polymerase activity of RNA replicases
US6369207B1 · kind B1 · utility
Assignee
Inventors
Key dates
| Filing date | Sep 14, 1999 |
| Grant date | Apr 9, 2002 |
| Priority date | — |
| Expiry date | Sep 14, 2019 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6853
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention entails methods, and kits for carrying them out, based on the discovery that an RNA replicase, such as Q&bgr; replicase, has DNA-dependent RNA polymerase (“DDRP”) activity with nucleic acid segments, including DNA segments and DNA:RNA chimeric segments, which comprise a 2′-deoxyribonucleotide or an analog thereof and which have sequences of RNAs that are autocatalytically replicatable by the replicase. The discovery of this DDRP activity provides methods of the invention for nucleic acid amplification wherein a nucleic acid, with a DNA segment with the sequence of an RNA that is autocatalytically replicatable by an RNA replicase, is provided as a substrate for the replicase. The replicase catalyzes synthesis, from the DNA segment, of the RNA, which the replicase then autocatalytically replicates. The invention entails use of the amplification methods in detecting nucleic acid analytes, as in nucleic acid probe hybridization assays. Such assays of the invention include those wherein a nucleic acid analyte is hybridized with one or more nucleic acid probes, which include or are processed to generate a DNA segment which is amplifiable through pr…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.