Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
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Evaluation of sperm DNA fragmentation using multiple methods: a comparison of their predictive power for male infertility

  • Received : 2018.10.26
  • Accepted : 2019.01.29
  • Published : 2019.03.31
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Abstract

Objective: The usual seminal profile has been customarily used for diagnosing male infertility based on an examination of semen samples. However, sperm DNA fragmentation has also been causally linked to reproductive failure, suggesting that it should be evaluated as part of male infertility assessments. To compare the ability of the five most widely utilized methodologies of measuring DNA fragmentation to predict male infertility and reactive oxygen species by Oxisperm kit assay. Methods: In this case-control study, which received ethical committee approval, the participants were divided into fertile and infertile groups (50 patients in each group). Results: The alkaline comet test showed the best ability to predict male infertility, followed by the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay, the sperm chromatin dispersion (SCD) test, and the sperm chromatin structure assay (SCSA), while the neutral comet test had no predictive power. For our patient population, the projected cut-off point for the DNA fragmentation index was 22.08% using the TUNEL assay, 19.90% using SCSA, 24.74% using the SCD test, 48.47% using the alkaline comet test, and 36.37% using the neutral comet test. Significant correlations were found between the results of the SCD test and those obtained using SCSA and TUNEL (r = 0.70 and r = 0.68, respectively; p< 0.001), and a statistically significant correlation was also found between the results of SCSA and the TUNEL assay (r = 0.77, p< 0.001). Likewise, the results of the alkaline comet test showed significant correlations with those of the SCD, SCSA, and TUNEL tests (r = 0.59, r = 0.57, and r = 0.72, respectively; p< 0.001). Conclusion: The TUNEL assay, SCSA, SCD, and the alkaline comet test were effective for distinguishing between fertile and infertile patients, and the alkaline comet test was the best predictor of male infertility.

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References

  1. Akashi T, Watanabe A, Komiya A, Fuse H. Evaluation of the sperm motility analyzer system (SMAS) for the assessment of sperm quality in infertile men. Syst Biol Reprod Med 2010;56:473-7. https://doi.org/10.3109/19396361003786293
  2. Practice Committee of the American Society for Reproductive Medicine. The clinical utility of sperm DNA integrity testing: a guideline. Fertil Steril 2013;99:673-7. https://doi.org/10.1016/j.fertnstert.2012.12.049
  3. Setti AS, Paes de Almeida Ferreira Braga D, Iaconelli A Jr, Aoki T, Borges E Jr. Twelve years of MSOME and IMSI: a review. Reprod Biomed Online 2013;27:338-52. https://doi.org/10.1016/j.rbmo.2013.06.011
  4. Perdrix A, Rives N. Motile sperm organelle morphology examination (MSOME) and sperm head vacuoles: state of the art in 2013. Hum Reprod Update 2013;19:527-41. https://doi.org/10.1093/humupd/dmt021
  5. Evenson DP, Jost LK, Marshall D, Zinaman MJ, Clegg E, Purvis K, et al. Utility of the sperm chromatin structure assay as a diagnostic and prognostic tool in the human fertility clinic. Hum Reprod 1999;14:1039-49. https://doi.org/10.1093/humrep/14.4.1039
  6. Sun JG, Jurisicova A, Casper RF. Detection of deoxyribonucleic acid fragmentation in human sperm: correlation with fertilization in vitro. Biol Reprod 1997;56:602-7. https://doi.org/10.1095/biolreprod56.3.602
  7. Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 1988;175:184-91. https://doi.org/10.1016/0014-4827(88)90265-0
  8. Fernandez JL, Muriel L, Rivero MT, Goyanes V, Vazquez R, Alvarez JG. The sperm chromatin dispersion test: a simple method for the determination of sperm DNA fragmentation. J Androl 2003;24:59-66. https://doi.org/10.1002/j.1939-4640.2003.tb02748.x
  9. Collins JA, Barnhart KT, Schlegel PN. Do sperm DNA integrity tests predict pregnancy with in vitro fertilization? Fertil Steril 2008;89:823-31. https://doi.org/10.1016/j.fertnstert.2007.04.055
  10. Wang YJ, Zhang RQ, Lin YJ, Zhang RG, Zhang WL. Relationship between varicocele and sperm DNA damage and the effect of varicocele repair: a meta-analysis. Reprod Biomed Online 2012;25:307-14. https://doi.org/10.1016/j.rbmo.2012.05.002
  11. Baehner RL, Boxer LA, Davis J. The biochemical basis of nitroblue tetrazolium reduction in normal human and chronic granulomatous disease polymorphonuclear leukocytes. Blood 1976;48:309-13. https://doi.org/10.1182/blood.V48.2.309.309
  12. Choi HS, Kim JW, Cha YN, Kim C. A quantitative nitroblue tetrazolium assay for determining intracellular superoxide anion production in phagocytic cells. J Immunoassay Immunochem 2006;27:31-44. https://doi.org/10.1080/15321810500403722
  13. Esfandiari N, Sharma RK, Saleh RA, Thomas AJ Jr, Agarwal A. Utility of the nitroblue tetrazolium reduction test for assessment of reactive oxygen species production by seminal leukocytes and spermatozoa. J Androl 2003;24:862-70. https://doi.org/10.1002/j.1939-4640.2003.tb03137.x
  14. Dominguez-Fandos D, Camejo MI, Ballesca JL, Oliva R. Human sperm DNA fragmentation: correlation of TUNEL results as assessed by flow cytometry and optical microscopy. Cytometry A 2007;71:1011-8. https://doi.org/10.1002/cyto.a.20484
  15. Evenson D, Gharagozloo P, Aitken RJ. Sperm chromatin structure assay ($SCSA^{(R)}$): the clinical utility of measuring sperm DNA damage and its potential improvement with supplemental antioxidants. JSM Invitro Fertil 2017;2:1008.
  16. Wang M, Sun J, Wang L, Gao X, Lu X, Wu Z, et al. Assessment of density gradient centrifugation (DGC) and sperm chromatin dispersion (SCD) measurements in couples with male factor infertility undergoing ICSI. J Assist Reprod Genet 2014;31:1655-63. https://doi.org/10.1007/s10815-014-0339-4
  17. Van Kooij RJ, de Boer P, De Vreeden-Elbertse JM, Ganga NA, Singh N, Te Velde ER. The neutral comet assay detects double strand DNA damage in selected and unselected human spermatozoa of normospermic donors. Int J Androl 2004;27:140-6. https://doi.org/10.1111/j.1365-2605.2004.00463.x
  18. Spano M, Bonde JP, Hjollund HI, Kolstad HA, Cordelli E, Leter G. Sperm chromatin damage impairs human fertility. The Danish First Pregnancy Planner Study Team. Fertil Steril 2000;73:43-50. https://doi.org/10.1016/S0015-0282(99)00462-8
  19. Brandriff B, Pedersen RA. Repair of the ultraviolet-irradiated male genome in fertilized mouse eggs. Science 1981;211:1431-3. https://doi.org/10.1126/science.7466400
  20. Evgeni E, Charalabopoulos K, Asimakopoulos B. Human sperm DNA fragmentation and its correlation with conventional semen parameters. J Reprod Infertil 2014;15:2-14.
  21. Lewis SE, John Aitken R, Conner SJ, Iuliis GD, Evenson DP, Henkel R, et al. The impact of sperm DNA damage in assisted conception and beyond: recent advances in diagnosis and treatment. Reprod Biomed Online 2013;27:325-37. https://doi.org/10.1016/j.rbmo.2013.06.014
  22. Chohan KR, Griffin JT, Lafromboise M, De Jonge CJ, Carrell DT. Comparison of chromatin assays for DNA fragmentation evaluation in human sperm. J Androl 2006;27:53-9. https://doi.org/10.2164/jandrol.05068
  23. Speyer BE, Pizzey AR, Ranieri M, Joshi R, Delhanty JD, Serhal P. Fall in implantation rates following ICSI with sperm with high DNA fragmentation. Hum Reprod 2010;25:1609-18. https://doi.org/10.1093/humrep/deq116
  24. Brahem S, Mehdi M, Landolsi H, Mougou S, Elghezal H, Saad A. Semen parameters and sperm DNA fragmentation as causes of recurrent pregnancy loss. Urology 2011;78:792-6. https://doi.org/10.1016/j.urology.2011.05.049
  25. Robinson L, Gallos ID, Conner SJ, Rajkhowa M, Miller D, Lewis S, et al. The effect of sperm DNA fragmentation on miscarriage rates: a systematic review and meta-analysis. Hum Reprod 2012;27:2908-17. https://doi.org/10.1093/humrep/des261
  26. Sharma RK, Sabanegh E, Mahfouz R, Gupta S, Thiyagarajan A, Agarwal A. TUNEL as a test for sperm DNA damage in the evaluation of male infertility. Urology 2010;76:1380-6. https://doi.org/10.1016/j.urology.2010.04.036
  27. Osman A, Alsomait H, Seshadri S, El-Toukhy T, Khalaf Y. The effect of sperm DNA fragmentation on live birth rate after IVF or ICSI: a systematic review and meta-analysis. Reprod Biomed Online 2015;30:120-7. https://doi.org/10.1016/j.rbmo.2014.10.018
  28. Simon L, Zini A, Dyachenko A, Ciampi A, Carrell DT. A systematic review and meta-analysis to determine the effect of sperm DNA damage on in vitro fertilization and intracytoplasmic sperm injection outcome. Asian J Androl 2017;19:80-90.
  29. Agarwal A, Majzoub A, Esteves SC, Ko E, Ramasamy R, Zini A. Clinical utility of sperm DNA fragmentation testing: practice recommendations based on clinical scenarios. Transl Androl Urol 2016;5:935-50. https://doi.org/10.21037/tau.2016.10.03

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