Terminal-phosphate-labeled nucleotides and methods of use
US7052839B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Apr 1, 2002 |
| Grant date | May 30, 2006 |
| Priority date | — |
| Expiry date | Aug 20, 2023 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6851
- WIPO fieldBasic materials chemistry
- WIPO sectorChemistry
Abstract
The present invention describes methods of detecting a nucleic acid in a sample, based on the use of terminal-phosphate-labeled nucleotides as substrates for nucleic acid polymerases. The methods provided by this invention utilize a nucleoside polyphosphate, dideoxynucleoside polyphosphate, or deoxynucleoside polyphosphate analogue which has a colorimetric dye, chemiluminescent, or fluorescent moiety, a mass tag or an electrochemical tag attached to the terminal-phosphate. When a nucleic acid polymerase uses this analogue as a substrate, an enzyme-activatable label would be present on the inorganic polyphosphate by-product of phosphoryl transfer. Cleavage of the polyphosphate product of phosphoryl transfer via phosphatase leads to a detectable change in the label attached thereon. When the polymerase assay is performed in the presence of a phosphatase, there is provided a convenient method for real-time monitoring of DNA or RNA synthesis and detection of a target nucleic acid.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.