[Molecular detection of Y chromosome microdeletions: a new approach based on the denaturing gradient gel...

Ten percent of couples trying for a child fail to conceive. In approximately 50% of cases, infertility is due to the inability of the male partner to produce spermatozoa in sufficient numbers to effect conception. Over the past 5 years, molecular studies have suggested that interstitial microdeletions in Yq11 represent an etiological factor for male infertility. Y-microdeletions have been detected in 12% of non-obstructive azoospermia and 6% of severe oligozoospermia. In general, microdeletions were detected by separate multiplex-polymerase chain reaction (PCR) reactions using primer pairs for single tagged sites (STSs) of all three azoospermia factor (AZF) regions (AZFa, AZFb and AZFc). This review describes the molecular methods and laboratory guidelines for molecular diagnosis of Y-chromosomal microdeletions. The diagnostics of Y-chromosomal microdeletions should be performed in two steps: in any case, the routine diagnostic should include six STS loci, two STS loci localised in each AZF region, and once a deletion is detected, the analysis can be extended to STS loci known to cross the proximal and the distal borderlines of each AZF region. Other molecular techniques such as DGGE, Southern blot should be performed to detect partial deletions of gene copies or mosaicism. These different molecular approaches should allow explaining 10% of male infertility, to evaluate the risk to pass the defect onto their male offspring (by intracytoplasmic sperm injection) and improve the genetic counselling of couples undergoing micromanipulative assisted reproduction.