Process for determining the quantity of a DNA fragment of interest by a method of enzymatic amplification of DNA

US5747246A · kind A · utility

Assignees

• Institut National de la Sante et de la Recherche Medicale (INSERM) · FR
• INSTITUT PASTEUR · FR

Inventors

• Christophe Pannetier · Paris, FR
• Madeleine Cochet · Fontenay-aux-Roses, FR
• Sylvie Darche · Brunoy, FR
• Philippe Kourilsky · Paris, FR

Key dates

• Filing date: Jun 21, 1994
• Grant date: May 5, 1998
• Expiry date: Jun 21, 2014

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6851
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention relates to a process for determining the quantity of a DNA fragment of interest in a sample wherein: PA1 1) a standard DNA fragment can be amplified with the same oligonucleotide primers is added to the sample to be analyzed containing the DNA fragment of interest, the standard DNA fragment and the fragment of interest differing in sequence and/or in size by not more than 10%, PA1 2) the DNA fragment of interest and the standard fragment are coamplified with the same primers, preferably to saturation of the amplification of the DNA fragment of interest, PA1 3) one or more labeled oligonucleotide primer(s), specific for the DNA fragment of interest and the standard fragment and different from the primers of step 2), is/are added to the reaction medium obtained in step 2), so that, after denaturation of the DNA, said primer(s) hybridize(s) with said fragments at a suitable site in order that an elongation with the DNA polymerase generates labeled DNA fragments of different sizes and/or sequences or with different labels according to whether they originate from the DNA fragment of interest or the standard fragment, respectively, and then PA1 4) the initial quanti…