Method for Quantitative Evaluation of a Rearrangement or a Targeted Genetic Recombination of an Individual and Uses Thereof

A method for the quantitative evaluation of a rearrangement or of a targeted genetic recombination of an individual, which method comprises at least:

• (a) the extraction of human genomic DNA from a biological sample,
• (b) the amplification of a segment of said genomic DNA, between a few hundred base pairs and several tens of kb in size, by multiplex PCR, in the presence:
• of one or more pairs of primers, selected so as to correspond to the following characteristics:
• at least one of said primers of one of said pairs of primers hybridizes upstream and/or at the 5′ end of a Vx gene to be amplified, which may be involved in said genetic rearrangement;
• at least the other of said primers of one of said pairs of primers hybridizes downstream and/or at the 3′ end of a Jy gene to be amplified, which may be involved in said genetic rearrangement;
• and of a DNA polymerase or a mixture of DNA polymerases for amplifying genomic DNA segments between a few hundred base pairs and several tens of kb in size, preferably greater than 10 kb in size, and having a correction activity that makes it possible to substantially improve the elongation;
• said amplification comprising, in addition to the initial denaturation step, cycles of denaturation, hybridization and elongation, in which the elongation steps are carried out at least for 10 minutes at 68° C.-72° C.;
• c) the separation of the gDNA fragments amplified, and
• d) the detection of the rearranged or recombined segments.