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Volume 13, Issue 6 (November 2011) 13, 877–880; 10.1038/aja.2011.51
Prevalent false positives of azoospermia factor a (AZFa) microdeletions caused by single-nucleotide polymorphism rs72609647 in the sY84 screening of male infertility
Qing Wu1, Guo-Wu Chen2, Tao-Fei Yan3, Hui Wang4, Yu-Ling Liu2, Zheng Li5, Shi-Wei Duan6, Fei Sun7, Yun Feng1 and Hui-Juan Shi4
1 Reproductive Medical Center, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
2 Shanghai Jiai Genetics and IVF Institute Center of China-USA, Shanghai 200011, China
3 Department of Obstetrics and Gynecology, Second Military Medical University, Shanghai 200433, China
4 China National Population and Family Planning Key Laboratory of Contraceptive Drugs and Devices, Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China
5 Shanghai Human Sperm Bank, Shanghai Institute of Andrology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200001, China
6 School of Medicine, Ningbo University, Ningbo 315033, China
7 Laboratory for Reproductive Biology, School of Life Science, University of Science and Technology of China, Hefei 230026, China
Correspondence: Dr Y Feng, (ivfruijin@yahoo.com.cn); Dr HJ Shi, (shihuijuan67@hotmail.com)
Received 20 December 2010; Revised 16 February 2011; Accepted 28 February 2011; Published online 18 July 2011
| Abstract |
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Multiplex polymerase chain reaction (PCR) has been widely used to detect Y-chromosome microdeletions, which is one of the major causes of male infertility. Both the European Academy of Andrology (EAA) and the European Molecular Genetics Quality Network (EMQN) have recommended the use of sY84 and sY86 markers for the detection of azoospermia factor a (AZFa) microdeletion during DNA testing for male infertility. In this study, a large-scale analysis of AZF microdeletion in a total of 630 Chinese males, including healthy semen donors (n=200), infertile males with normal sperm count (n=226) and patients with either nonobstructive azoospermia or severe oligozoospermia (n=204), was performed. A series of nine sequence-tagged site (STS) markers from the AZF region of the Y chromosome was used to detect microdeletions. All primers were designed based on the recommendations of the National Center for Biotechnology Information. An unusually high incidence (73/630, 11.6%) of sY84-absent but sY86-present genotypes was observed in the AZFa microdeletion screening. Sequencing the sY84-flanking region revealed a total of 73 patients with sY84-absent but sY86-present genotypes have a T-to-G transversion at the fifth base from the 5' end of the reverse sY84 primer. These prevalent false positives, which were not only observed in infertile men, but also observed in donors, resulted from a single-nucleotide polymorphism (SNP) named rs72609647 in the targeting sequence of the reverse sY84 primer. Our study suggests that a pre-screening of existence of rs72609647 polymorphism can prevent the frequent false positive results of AZFa microdeletions detection in the infertile Chinese males. Given the SNP rs72609647 was recently found in a deep sequencing of a Chinese individual, the current EAA and EMQN standards may need to be scrutinized among different populations to avoid the potential genetic variations in the primer binding sequences.
Keywords: male infertility; multiplex polymerase chain reaction; rs72609647; single-nucleotide polymorphism; sY84; Y-chromosome microdeletion
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