Extremely Sensitive, Background-Free Gene Detection Using Binary Probes and Qβ Replicase
Abstract
We have developed a specific and sensitive nucleic acid amplification assay that is suitable for routine gene detection. The assay is based on a novel molecular genetic strategy in which two different RNA probes are hybridized to adjacent positions on a target nucleic acid and then ligated to form an amplifiable reporter RNA. The reporter RNA is then replicated up to a hundred billion-fold in a 30-min isothermal reaction that signals the presence of the target. The assay can detect fewer than 100 nucleic acid molecules; it provides quantitative results over a wide range of target concentrations and it employs a universal format that can detect any infectious agent.
- Publication:
-
Proceedings of the National Academy of Science
- Pub Date:
- May 1996
- DOI:
- 10.1073/pnas.93.11.5395
- Bibcode:
- 1996PNAS...93.5395T