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A novel universal multiplex PCR improves detection of AZFc Y-chromosome microdeletions

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Abstract

Purpose

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.

Methods

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.

Results

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).

Conclusions

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).

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Acknowledgments

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).

Author contributions

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.

Author information

Authors and Affiliations

  1. Department of Clinical Laboratory, Renmin Hospital of Wuhan University, Wuhan, Hubei, People’s Republic of China

    Hong-Yun Zheng, Yan Li & Yong-Qing Tong

  2. Clinical Molecular Diagnostic Center, Renmin Hospital of Wuhan University, Wuhan, Hubei, People’s Republic of China

    Hong-Yun Zheng, Yan Li & Yong-Qing Tong

  3. Department of Gynecology, Renmin Hospital of Wuhan University, Wuhan, Hubei, People’s Republic of China

    Fu-Jin Shen

Authors
  1. Hong-Yun Zheng
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  2. Yan Li
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Corresponding author

Correspondence to Yan Li.

Additional information

Capsule Traditional multiplex PCR (M-PCR) has been widely used to detect Y-chromosome microdeletions, however, its specificity is not very high. In the present study, we established a novel universal primer-multiplex-PCR (UP-M-PCR) method to overcome the disadvantage. On the other hand, we detected 15 STS to find all the possible deletion sites, which is more accurate for diagnosis of male infertility compared to the past used 6 STS.

In the current manuscript, we eatablished 4 stable and reliable UP-M-PCR reaction systems to detected the 15 sequence-tagged sites (STS) for the AZFa, AZFb, AZFc and AZFd regions of the Y-chromosome. 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). However, there were only 0.5 % and 1 % of AZFb and AZFc ’s respectively by traditional M-PCR. Thus data showed the great advantages of UP-M-PCR, which could provide a more accurate evidence for the diagnosis of these patients with severe oligozoospermia or azoospermia.

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Zheng, HY., Li, Y., Shen, FJ. et al. A novel universal multiplex PCR improves detection of AZFc Y-chromosome microdeletions. J Assist Reprod Genet 31, 613–620 (2014). https://doi.org/10.1007/s10815-014-0204-5

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  • DOI: https://doi.org/10.1007/s10815-014-0204-5

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