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Actas Urol Esp. 2014 Nov;38(9):608-12. doi: 10.1016/j.acuro.2014.02.022. Epub 2014 Jun 2.

Fragmentation of sperm DNA using the TUNEL method.

[Article in English, Spanish]

Author information

  • 1Departamento de Bioquímica Clínica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Hospital de Clínicas José de San Martín, Buenos Aires, Argentina.
  • 2Departamento de Bioquímica Clínica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Hospital de Clínicas José de San Martín, Buenos Aires, Argentina. Electronic address: gmendeluk@ffyb.uba.ar.

Abstract

OBJECTIVES:

To establish the validity of the TUNEL assay in determining sperm DNA fragmentation, the relationship between the degree of fragmentation and the seminal parameters and the sample needed to conduct the test.

MATERIAL AND METHODS:

We used semen samples from healthy fertile men (n=33), patients who consulted for infertility with a prescription for the TUNEL assay (n=77) and patients with intracytoplasmic sperm injection failure (n=20), analyzed according to the 2010 WHO. The TUNEL/propidium iodide test was performed by flow cytometry, on baseline and post-swim-up samples.

RESULTS:

The cutoff value for the TUNEL assay (ROC curves) was 26%, with a sensitivity and specificity of 85% and 89%, respectively. The pre-swim-up and post-swim-up medians of the results from the TUNEL assay showed no significant differences (17.0% vs. 12.9%, respectively). However, 39.1% of the samples showed a difference greater than 15 in absolute value between the results of the baseline and post-swim-up TUNEL assays. The linear correlation study of the morphology, mobility and vitality using the post-swim-up TUNEL assay showed a greater correlation than preselection, with significant results (r: -0.394, P<.0001; r: -0.461, P<.0001; r: -0.526, P<.0001).

CONCLUSIONS:

The TUNEL assay is a valid test for clinical use. DNA fragmentation is a factor independent from traditional semen tests. We found a greater susceptibility to damage generated in the laboratory procedures in the samples with lower quality. The sample of choice for evaluating DNA fragmentation will depend on whether the clinician is treating a natural or assisted fertilization.

Copyright © 2014 AEU. Published by Elsevier Espana. All rights reserved.

KEYWORDS:

Fragmentación del núcleo espermático; Parámetros espermáticos; Sperm nucleus fragmentation; Sperm parameters; Susceptibilidad al daño en el laboratorio; Susceptibility to damage in the laboratory; Swim up; TUNEL; Test validity; Validez de la prueba

PMID:
24889159
[PubMed - in process]
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