Oligonucleotide modification, signal amplification, and nucleic acid detection by target-catalyzed product formation
US6110677A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Jan 30, 1998 |
| Grant date | Aug 29, 2000 |
| Priority date | — |
| Expiry date | Jan 30, 2018 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6823
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method is disclosed for modifying an oligonucleotide, which method has application to the detection of a polynucleotide analyte. An oligonucleotide is reversibly hybridized with a polynucleotide, for example, a polynucleotide analyte, in the presence of a 5'-nuclease under isothermal conditions. The polynucleotide analyte serves as a recognition element to enable a 5'-nuclease to cleave the oligonucleotide to provide (i) a first fragment that is substantially non-hybridizable to the polynucleotide analyte and (ii) a second fragment that lies 3' of the first fragment (in the intact oligonucleotide) and is substantially hybridizable to the polynucleotide analyte. At least a 100-fold molar excess of the first fragment and/or the second fragment are obtained relative to the molar amount of the polynucleotide analyte. The presence of the first fragment and/or the second fragment is detected, the presence thereof indicating the presence of the polynucleotide analyte. The method has particular application to the detection of a polynucleotide analyte such as DNA. Kits for conducting methods in accordance with the present invention are also disclosed.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.