Nucleic acid amplification employing ligatable hairpin probe and transcription
US5215899A · kind A · utility
Assignee
Inventor
Key dates
| Filing date | Aug 23, 1990 |
| Grant date | Jun 1, 1993 |
| Priority date | — |
| Expiry date | Aug 23, 2010 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY10S436/815
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Specific nucleic acid sequences are amplified through the use of a hairpin probe which, upon hybridization with and ligation to, a target sequence is capable of being transcribed. The probe comprises a single stranded self-complementary sequence which, under hybridizing conditions, forms a hairpin structure having a functional promoter region, and further comprises a single stranded probe sequence extending from the 3' end of the hairpin sequence. Upon hybridization with a target sequence complementary to the probe sequence and ligation of the 3' end of the hybridized target sequence to the 5' end of the hairpin probe, the target sequence is rendered transcribable in the presence of a suitable RNA polymerase and appropriate ribonucleoside triphosphate (rNTPs). Amplification is accomplished by hybridizing the desired target nucleic acid sequence with the probe, ligating the target sequence to the probe, adding the RNA polymerase and rNTPs to the separated hybrids, and allowing transcription to proceed until a desired amount of RNA transcription product has accumulated. The amplification method is particularly useful in assays for the detection of particular nucleic acid sequences.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.