A novel universal multiplex PCR improves detection of AZFc Y-chromosome microdeletions
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To determine the frequencies and the characteristics of Y chromosome microdeletions (pl) in infertile men from central China to perform appropriate therapeutic choices by updated multiplex-PCR.
In this study, we established a novel universal primer-multiplex-PCR (U-M-PCR) method to overcome the disadvantages of traditional multiplex PCR (M-PCR). We chose 15 sequence-tagged sites (STS) for detection of Y chromosome microdeletions. 540 infertile male patients and 100 healthy male controls were selected in the study.
Of the 540 male infertility patients, 48 Y-chromosome microdeletions were detected, with a total deletion rate of 8.9 %. Of these deletions, the rate of AZFa deletions (sY84) was 0.5 % (3/540), the rate of AZFb deletions (sY143) was 0.7 % (4/540) and the rate of AZFc deletions (sY242, sY254 and sY255) was 7.6 % (41/540). Compared with AZF deletion rates by M-PCR, we found U-M-PCR could detect AZFc deletion more specifically (1.0 % & 7.6 %). No Y-chromosome microdeletions were detected in the 100 males with normal semen (the control group).
U-M-PCR method was more specific to detect AZFc microdeletions. It is necessary to use the U-M-PCR method to offer genetic screening and counseling to infertile men prior to intracytoplasmic sperm injection (ICSI) or in-vitro fertilization (IVF).
KeywordsMale infertility Y-chromosome Microdeletions Multiplex PCR AZFc
This research supported by the forty-ninth issue of the China Postdoctoral Science Foundation, first-class funding (Grant No. 20110490119), National Natural Science Foundation of China (Grant No. 81100959) and National Clinical Key Specialty Construction Projects of the Health Ministry (2010).
ZHY carried out the multiplex PCR, performed the statistical analysis and drafted the manuscript. SFJ and TYQ collected the samples and extracted genomic DNA from peripheral blood. LY designed the study and helped to draft the manuscript. All authors read and approved the final manuscript.
Competing financial interests
The authors have nothing to disclose.
- 3.Efstratiou A, George RC. Laboratory guidelines for the diagnosis of infections caused by Corynebacterium diphtheriae and C. ulcerans. World Health Organization. Commun Dis Public Health/PHLS. 1999;2(4):250–7.Google Scholar
- 8.Cai ZM. [Y chromosome microdeletion and male infertility: past, present and future]. Zhonghua nan ke xue. Natl J Androl. 2010;16(5):387–94.Google Scholar
- 12.Silber SJ, Disteche CM. In: Pagon RA, Adam MP, Bird TD, Dolan CR, Fong CT, Stephens K, editors. Y chromosome infertility. Seattle: GeneReviews; 1993.Google Scholar
- 24.Mirfakhraie R, Mirzajani F, Kalantar SM, Montazeri M, Salsabili N, Pourmand GR, et al. High prevalence of AZFb microdeletion in Iranian patients with idiopathic non-obstructive azoospermia. Indian J Med Res. 2011;132:265–70.Google Scholar
- 26.Bu Y, Huang H, Wu HP, Zhang XD, Zhou GH, Cui YX, et al. Direct multiplex-PCR from whole blood for rapid detection of Y chromosome microdeletions. Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi. Chin J Med Genet. 2008;25(4):406–9.Google Scholar