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Fragmentation of sperm DNA using the TUNEL method.

P H Chenlo1, S M Curi1, M N Pugliese1

  • 1Departamento de Bioquímica Clínica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Hospital de Clínicas José de San Martín, Buenos Aires, Argentina.

Actas Urologicas Espanolas
|June 4, 2014
PubMed
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The TUNEL assay is a valid clinical tool for assessing sperm DNA fragmentation, an indicator independent of traditional semen parameters. Sample choice for testing depends on fertilization method.

Area of Science:

  • Reproductive Biology
  • Andrology
  • Clinical Diagnostics

Background:

  • Sperm DNA fragmentation is a critical factor affecting male fertility.
  • The Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay is a potential method for its evaluation.
  • Establishing the assay's clinical validity and optimal sample selection is crucial.

Purpose of the Study:

  • To validate the TUNEL assay for sperm DNA fragmentation assessment.
  • To investigate the relationship between DNA fragmentation and seminal parameters.
  • To determine the optimal sample type for TUNEL testing.

Main Methods:

  • Semen samples from fertile men, infertile patients, and ICSI failure cases were analyzed.
  • The TUNEL/propidium iodide test was performed using flow cytometry.
Keywords:
Fragmentación del núcleo espermáticoParámetros espermáticosSperm nucleus fragmentationSperm parametersSusceptibilidad al daño en el laboratorioSusceptibility to damage in the laboratorySwim upTUNELTest validityValidez de la prueba

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  • Baseline and post-swim-up samples were evaluated.

    • Main Results:

    • The TUNEL assay demonstrated a cutoff value of 26% with 85% sensitivity and 89% specificity.
    • No significant difference was observed between pre-swim-up and post-swim-up TUNEL results.
    • Sperm DNA fragmentation showed significant negative correlations with morphology, motility, and vitality post-swim-up.

    Conclusions:

    • The TUNEL assay is validated for clinical application in assessing sperm DNA fragmentation.
    • Sperm DNA fragmentation is an independent factor from conventional semen analysis parameters.
    • Sample selection for TUNEL assay should consider the intended fertilization approach (natural vs. assisted).