Nucleic acid amplification method hybridization signal amplification method (HSAM)

USRE38442E1 · kind E1 · reissue

Assignee

• Mount Sinai School of Medicine · US

Inventors

• David Zhang · Elmont, US
• Margaret Brandwein · Elmont, US

Key dates

• Filing date: Mar 2, 2001
• Grant date: Feb 24, 2004
• Expiry date: Mar 2, 2021

Classification

• Technology area (CPC —): General

Abstract

An improved method allowing for rapid sensitive and standardized detection of a target nucleic acid from a pathogenic microorganism or virus or normal or abnormal gene in a sample is provided. The method involves hybridizing a target nucleic acid to several non-overlapping oligonucleotide probes that hybridize to adjacent regions in the target nucleic acid, the probes being referred to capture/amplification probes and amplifications probes, respectively, in the presence of paramagnetic beads coated with a ligand binding moiety. Through the binding of a ligand attached to one end of the capture/amplification probe and the specific hybridization of portions of the probes to adjacent sequences in the target nucleic acid, a complex comprising the target nucleic acid, the probes and the paramagnetic beads is formed. The probes may then ligated together to form a contiguous ligated amplification sequence bound to the beads, which complex may be denatured to remove the target nucleic acid and unligated probes. Alternatively, separate capture and amplification probes may be used which form continuous full-length or circular probes, and may be directly detected or amplified using a suitable…