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Quantitative PCR technique for the identification of microrearrangements of the AZFc region
Virginie Rozé1, Jean Luc Bresson, Florence Fellmann
1EA MENRT 3185 Génétique et Reproduction - IFR133 IBCT, Faculté de Médecine et de Pharmacie, Besançon, France. virginie.roze@univ-fcomte.fr
Purpose:
The AZFc region spans about 3.5 Mb and contains many amplicons causing recombination events. Several papers have reported the occurrence of AZFc partial deletions resulting from non allelic homologous recombination (NAHR) ("gr-gr", "b1-b3" or "b2-b3" deletions), particularly in infertile patients. DAZ genes are present in 4 copies and rearrangements involve a modification of the number of DAZ genes.
Methods:
In addition to STS plus/minus PCR, we developed a quantitative technique using real time PCR (Q-PCR) to determine the number of DAZ genes. Fourteen DNA controls were selected to validate the use of Q-PCR to detect AZFc microrearrangements, and sperm DNA samples from 30 fertile men were studied.
Results:
Rearrangements of 14 controls were well identified with Q-PCR, and 2 AZFc partial deletions were detected in fertile men (1 "gr-gr" and 1 "b2-b3").
Conclusion:
Q-PCR represents a well-adapted method to detect microrearrangements of the Y-chromosome, complementary to STS analysis.
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