Screening for Y chromosome azoospermia factor region microdeletions using modified multiplex PCR

Abstract

Objective To establish an universal primer-multiplex PCR system for diagnosis of Y chromosome AZF region microdeletions in 262 patients with non-obstructive azoospermic and severe oligozoospermic male infertility. Methods In each panel of multiplex PCR, YUP and YCP containing a fragment of non-human DNA sequence at their 5' ends were designed. The universal primers and chimiric primers were employed for the amplification at the same multiplex PCR system to screen for the Y chromosome AZF region ( a, b and c) microdeletions in 262 non-obstructive azoospermic and severe oligozoospermic male infertility patients. Results Thirty-three out of 262 patients (12. 60% ) were detected with Y chromosome AZF microdeletions. Among them, 27 cases were AZF c microdeletions and 6 ones were AZF b + c microdeletions. These results were in agreement with the results from EMQN method. There was no false-positivity. The gel electrophoresis for detection of multiple STS from both methods showed that the sY84,sY86, sY127, sY134, sY254, sY255, SRY bands were homogeneous and clear with similar brightness. Conclusion The modified multiplex PCR is suitable for screening of Y chromosome AZF microdeletions in non-obstructive azoospermic and severe digozoospermic male infertility patients. Key words: Infertility,male; Y chromosome; Chromosome deletion; Polymerase chain reaction