Bioluminescence regenerative cycle (BRC) for nucleic acid quantification
US7141370B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jun 28, 2002 |
| Grant date | Nov 28, 2006 |
| Priority date | — |
| Expiry date | Jun 28, 2022 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/689
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention concerns methods of quantifying nucleic acids using a bioluminescence regenerative cycle (BRC). In BRC, steady state levels of bioluminescence result from processes that produce pyrophosphate. Pyrophosphate reacts with APS in the presence of ATP sulfurylase to produce ATP. The ATP reacts with luciferin in a luciferase-catalyzed reaction, producing light and regenerating pyrophosphate. The pyrophosphate is recycled to produce ATP and the regenerative cycle continues. Because the kinetic properties of ATP sulfurylase are much faster than luciferase, a steady state results wherein concentrations of ATP and pyrophosphate and the rate of light production remain relatively constant. Photons are counted over a time interval to determine the number of target molecules present in the initial sample. The BRC process has a controllable dynamic range up to seven orders of magnitude and is sensitive enough to detect a few thousand molecules of target nucleic acid.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.