[Detection of Y chromosome microdeletions in semen of patients with azoospermia: study of 241 cases].
OBJECTIVE
To set up a simple and reliable method to screen Y chromosome microdeletions in semen of azoospermic patients, and to explore the incidence and loci of Y chromosome microdeletions in Chinese azoospermia.


METHODS
Two hundred and forty-one semen samples, 51 containing blood, were collected from 241 Chinese azoospermic patients. 45 normal semen samples and 1 anticoagulated blood sample from female were used as controls. DNA was quickly abstracted by incubating the cells with a lysis buffer containing polymerase chain reaction (PCR) buffer and protease K, and was used to detect the deletion of 15 kinds of sequence tagged site (STS) distributed in AZFa, AZFb, and AZFc by 4 sets of multiplex PCR. Agarose electrophoresis was used to observe and compare the PCR products from the semen samples and their corresponding blood samples, and the PCR products from 1 semen sample were confirmed by sequencing.


RESULTS
All multiplex PCR reactions were amplified successfully. The sequencing of the PCR products from the semen samples confirmed the PCR reactions. No microdeletion was detected in the 45 normal semen samples. Microdeletion was found in 26 out of the 241 semen samples of azoospermic patients (10.8%): 2 patients (7.7%) had the deletions located in AZFa, 2 patients (7.7%) in AZEb, 3 patients (11.5%) in both AZFb + AZFc, and other 19 patients (73.1%) in AZFc. The detection results of the blood samples were completely consistent with those of the semen samples. The STS deletion recommended by European Academy of Andrology (EAA) and European Molecular Genetics Quality Network (EMQN) were all found in these 26 cases.


CONCLUSIONS
The semen samples of azoospermic patients present a convenient, reliable, and noninvasive substitute for blood in screening of Y chromosome microdeletions, and can be employed in study and clinical examination. The EAA/EMQN recommendations allow the detection of complete AZF deletions in Chinese azoospermia.
Li
Hong-gang
HG
Family Planning Research Institute, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Ding
Xiao-fang
XF
Wang
Cheng
C
Zhao
Jiang-xia
JX
Zuo
Ming-da
MD
Xiong
Cheng-liang
CL
D053713:Azoospermia / Q000175:diagnosis / Q000235:genetics*; D002872:Chromosome Deletion; D041322:Chromosomes, Human, Y / Q000235:genetics*; D005260:Female; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D007248:Infertility, Male / Q000235:genetics; D008297:Male; D016133:Polymerase Chain Reaction; D012661:Semen / Q000378:metabolism*