[Screening of azoospermia factor microdeletions on Y chromosome in infertile men by QF-PCR]

To assess the application of quantitative fluorescent polymerase chain reaction (QF-PCR) on rapid screening of azoospermia factor (AZF) microdeletions, 1218 infertile men with non-obstructive azoospermia or oligospermia were detected for 9 sequence tagged sites (STSs) in AZF region by multiplex QF-PCR combined with capillary electrophoresis. AMEL (amelogenin) as well as SRY (sex-determining region of Y chromosome) located on short arm of sex chromosome was selected as internal control. Karyotyping was performed on Giemsa-banded metaphase chromosomes of peripheral blood lymphocytes. Of the 1218 patients, 105 (8.62%) were identified as AZF microdeletions. Deletion of AZFc (67.62%) was the most frequent, followed by deletion of AZFb,c (20.95%), AZFb (7.62%) and AZFa (3.81%). Five patients presented with deletions of both AZFa,b,c and AMEL-Y, indicating sex reversal which was confirmed to be 46,XX by karyotyping. Among the 105 patients with AZF microdeletions, 16 were karyotyped as chromosomal anomalies, most commonly 46,XY,Yqh- (75%, 12/16). In addition, of the total 1218 patients examined, 86 patients showed abnormal AMEL-X/AMEL- Y ratio, suggesting a possibility of sex chromosome anomalies, and 68 of them were verified as sex chromosome aneuploid by karyotyping. Multiplex QF-PCR is capable to detect all markers in one reaction and is also suggestive for sex chromosome anomalies. It could serve as an effective technique for screening Y-microdeletions, and thus have general application in diagnosis and treatment of male infertility.