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Molecular analysis of testis biopsy and semen pellet as complementary methods with histopathological analysis of testis in non-obstructive azoospermia

  • Gonadal Physiology and Disease
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

Non-obstructive azoospermia (NOA) is one the many causes of male infertility (10 %) resulting from testicular failure. Multiple testicular biopsies fail to find mature sperm in at least 50 % of cases Therefore; hunting for sensitive and specific biomarkers of spermatogenesis that could better determine the fertility status in NOA can lead to improved management of male infertility. Therefore, we evaluated sperm production through analyses of germ cell-specific transcripts (DAZ, TSPY1, SPTRX3 and SPTRX1) in semen and testicular biopsies of men with azoospermia.

Methods

We collected semen (N = 83) and testis biopsies (N = 31) from men with non-obstructive azoospermia. We later extracted RNA and synthesized cDNA using washed semen precipitate and testicular tissues. We also performed semi-nested PCR with designed specific primers. Using H&E method, an expert pathologist performed the histopathological evaluation. Having categorized the patients into three groups based on histopathological results, we calculated the agreement between molecular results of semen and tissues with histopathological findings for each patient using Kappa statistical test.

Results

Molecular findings of precipitated semen and testicular tissues were in disagreement with histopathological results in most cases. Molecular analysis of testis biopsies showed significant difference (Kappa coefficient = 0.009, P value = 0.894) with histopathological results; TSPY1, DAZ, SPTRX3 and SPTRX1 were respectively detected in 94 %, 94 %, 17.6 % and 52.9 % of men diagnosed with germ cell aplasia.

Conclusions

Molecular analysis of semen does not provide sufficient sensitivity and specificity to be used as a screening test at the present time, but it is a useful adjunct to histopathological methods in men with NOA. Spermatid/sperm specific transcripts indicated the possibility to find mature sperm following repeated multiple testicular sperm extraction (TESE) or microdisection TESE (mTESE).

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Acknowledgments

This study was financially supported by grant No. 1424-33 from the Academic Center for Education, Culture and Research (ACECR) in Tehran, Iran. We would like to thank Ms. Elham Savadi Shirazi for helping with the sample collection and Mr. Iman Derakhsh for the artwork preparation.

Author information

Authors and Affiliations

  1. Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Poursina Ave, 16 Azar St. Keshavarz BLVD, Tehran, Iran, 1417613151

    Maryam Eghbali & Mohammad Hossein Modarressi

  2. Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, P.O.Box:19615-1177, Tehran, Iran

    Mohammad Reza Sadeghi

  3. Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran

    Niknam Lakpour

  4. Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran

    Hale Edalatkhah & Mohammad Mehdi Akhondi

  5. Epidemiology and Biostatistics Department, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

    Hojjat Zeraati

  6. Avicenna Infertility Clinic, Avicenna Research Institute, ACECR, Tehran, Iran

    Haleh Soltanghoraee

  7. Iranian Tissue Bank Research and Preparation Center, Tehran University of Medical Sciences, Tehran, Iran

    S. Behnam Hashemi

  8. Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Poursina Ave, 16 Azar St. Keshavarz BLVD, Tehran, Iran, 14176-13151

    Mohammad Hossein Modarressi

Authors
  1. Maryam Eghbali
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  2. Mohammad Reza Sadeghi
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  3. Niknam Lakpour
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  4. Hale Edalatkhah
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  5. Hojjat Zeraati
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  6. Haleh Soltanghoraee
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  7. Mohammad Mehdi Akhondi
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  8. S. Behnam Hashemi
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  9. Mohammad Hossein Modarressi
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Corresponding authors

Correspondence to Mohammad Reza Sadeghi or Mohammad Hossein Modarressi.

Additional information

Capsule The molecular analysis of testicular tissues and semen precipitates for the presence of germ cell-specific transcripts is a diagnostic test that could be of help in the detection of active spermatogenesis in men with non-obstructive azoospermia.

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Eghbali, M., Sadeghi, M.R., Lakpour, N. et al. Molecular analysis of testis biopsy and semen pellet as complementary methods with histopathological analysis of testis in non-obstructive azoospermia. J Assist Reprod Genet 31, 707–715 (2014). https://doi.org/10.1007/s10815-014-0220-5

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  • DOI: https://doi.org/10.1007/s10815-014-0220-5

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