Mutation analysis using mass spectrometry

US6503710B2 · kind B2 · utility

Inventors

• Ivo Glynne Gut · Paris, FR
• Kurt Berlin · Stahnsdorf, DE
• Doris Lechner · Paris, FR
• Hans Lehrach · Berlin, DE

Key dates

• Filing date: May 27, 1999
• Grant date: Jan 7, 2003
• Expiry date: May 27, 2019

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y10T436/25125
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The invention presents a method for examining genetic material (deoxyribonucleic acid, DNA) to detect the presence of pre-known mutations, especially single nucleotide polymorphisms (SNP), using mass spectrometry with ionization by matrix-assisted laser desorption (MALDI). The invention uses nucleoside triphosphates with modified sites for the method of primer extension in a duplicating, enzymatic reaction and at least partially removal of primers from the extension product, in combination with product neutralization by chemical treatment of the modified sites, so that the resulting DNA products can be, by using special matrix materials, preferredly ionized in an adduct-free form over other constituents in the reaction solution without any further cleaning. The method is particularly suitable for simultaneous identification of several mutations by multiplexing.