Detection of nucleic acid sequence differences using the ligase detection reaction with addressable arrays

US7892747B2 · kind B2 · utility

Assignee

• Cornell Research Foundation, Inc. · US

Inventors

• Francis Barany · Baltimore, US
• George Barany · Falcon Heights, US
• Robert P. Hammer · Baton Rouge, US
• Maria Kempe · Lund, SE
• Herman Blok · Wemeldinge, NL
• Monib Zirvi · Monmouth Junction, US

Key dates

• Filing date: Mar 16, 2010
• Grant date: Feb 22, 2011
• Expiry date: Mar 16, 2030

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C40B60/14
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention describes a method for identifying one or more of a plurality of sequences differing by one or more single base changes, insertions, deletions, or translocations in a plurality of target nucleotide sequences. The method includes a ligation phase, a capture phase, and a detection phase. The ligation phase utilizes a ligation detection reaction between one oligonucleotide probe, which has a target sequence-specific portion and an addressable array-specific portion, and a second oligonucleotide probe, having a target sequence-specific portion and a detectable label. After the ligation phase, the capture phase is carried out by hybridizing the ligated oligonucleotide probes to a solid support with an array of immobilized capture oligonucleotides at least some of which are complementary to the addressable array-specific portion. Following completion of the capture phase, a detection phase is carried out to detect the labels of ligated oligonucleotide probes hybridized to the solid support.