Method for studying v(d)j combinatory diversity

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2706667

The invention relates to a method for analysing the diversity of the catalogue of T and/or B lymphocytes in an individual, based on the amplification, from a sample, of genomic DNA fragments by PCR multi-n-plexes, with n=2, carried out with a combination of at least 3 primers defining at least 2 primer couples, each of which includes a primer specifically hybridising upstream and/or in a given V or D gene and a primer specifically hybridising downstream and/or in a given J gene, in order to obtain the amplification of at least two fragments characteristic of two distinct V-J or D-J rearrangements from each primer couple. The invention also relates to the applications of this method, in particular in the treatment follow-up or in the diagnosis and/or prognosis of certain diseases.

L'invention porte sur une méthode d'analyse de la diversité du répertoire des lymphocytes T et/ou B d'un individu, basée sur l'amplification, à partir d'un échantillon, de fragments d'ADN génomique par des PCR multi-n-plexes avec n>=2, effectuées avec une combinaison d'au moins 3 amorces, constituant au moins 2 couples d'amorces dont chacun est constitué d'une amorce s'hybridant spécifiquement en amont et/ou dans un gène V ou D donné et d'une amorce s'hybridant spécifiquement en aval et/ou dans un gène J donné, de façon à permettre l'amplification d'au moins deux fragments caractéristiques de deux réarrangements V-J ou D-J distincts, à partir de chaque couple d'amorces. L'invention concerne également les applications de cette méthode, notamment dans le suivi d'un traitement ou dans le diagnostic et/ou le pronostic de certaines pathologies.

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