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Nat Biotechnol. 1999 Aug;17(8):804-7.

Detection of PCR products using self-probing amplicons and fluorescence.

Author information

1
AstraZeneca Diagnostics, Gadbrook Park, Rudheath, Northwich, Cheshire CW9 7RA, UK. david.whitcombe@diagnostics.zeneca.com

Abstract

Molecular diagnostics is progressing from low-throughput, heterogeneous, mostly manual technologies to higher throughput, closed-tube, and automated methods. Fluorescence is the favored signaling technology for such assays, and a number of techniques rely on energy transfer between a fluorophore and a proximal quencher molecule. In these methods, dual-labeled probes hybridize to an amplicon and changes in the quenching of the fluorophore are detected. We describe a new technology that is simple to use, gives highly specific information, and avoids the major difficulties of the alternative methods. It uses a primer with an integral tail that is used to probe an extension product of the primer. The probing of a target sequence is thereby converted into a unimolecular event, which has substantial benefits in terms of kinetics, thermodynamics, assay design, and probe reliability.

PMID:
10429248
DOI:
10.1038/11751
[Indexed for MEDLINE]
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