Abstract: Objective To evaluate the feasibility of application of real-time PCR (RT-PCR) method in Y chromosome microdeletions in male infertility. Methods A total of 443 male infertility patients in our hospital from March to October in 2015 were recruited in this study. Eight sequence tagged sites (STS) in 4 regions (AZFa, AZFb, AZFc and AZFd) of Y chromosome weredetected by RT-PCR. Moreover, samples with Y chromosome microdeletions were confirmed by multiplex PCR-electrophoresis. Results Three cases of Y chromosome microdeletions were identified and confirmed by RT-PCR. They haddefections as following: 2 AZFc+d and 1 AZFb+c+d, totally accounting for 10.3% of non-obstructive azoospermia cases. Conclusion Real-time PCR method allows a rapid and simple screening for Y chromosome microdeletions in male infertility, which is of clinical value for screening patients with infertility.