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TUNEL assay-Standardized method for testing sperm DNA fragmentation.

Rakesh Sharma1, Concetta Iovine1, Ashok Agarwal1

  • 1American Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH, USA.

Andrologia
|July 25, 2020
PubMed
Summary
This summary is machine-generated.

Sperm DNA fragmentation (SDF) impacts male fertility. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labelling (TUNEL) assay is a reliable method for assessing DNA breaks and male infertility.

Keywords:
DNA fragmentationTUNEL assayflow cytometermale infertilitysperm

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Area of Science:

  • Reproductive biology
  • Molecular diagnostics
  • Andrology

Background:

  • Sperm DNA integrity is crucial for successful reproduction, including fertilization, implantation, and fetal development.
  • Sperm DNA fragmentation (SDF) is frequently observed in infertile men and is linked to diminished reproductive outcomes.
  • Various methods exist to assess SDF, but the TUNEL assay is gaining prominence.

Purpose of the Study:

  • To review the protocol, quality control, and measurement of sperm DNA fragmentation using the TUNEL assay with flow cytometry.
  • To discuss factors influencing TUNEL assay results.
  • To explore the clinical implications of the TUNEL assay in evaluating male infertility.

Main Methods:

  • Detailed description of the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labelling (TUNEL) assay protocol.
  • Utilization of a benchtop flow cytometer for quantitative measurement of sperm DNA fragmentation.
  • Emphasis on quality control measures for accurate SDF assessment.

Main Results:

  • The TUNEL assay directly quantifies both single- and double-strand DNA breaks in sperm.
  • The review outlines a standardized approach to measuring SDF using flow cytometry.
  • Factors affecting TUNEL assay results and potential challenges are discussed.

Conclusions:

  • The TUNEL assay is a valuable and increasingly popular method for assessing sperm DNA fragmentation.
  • Accurate measurement of SDF using TUNEL can aid in diagnosing male infertility.
  • Understanding the factors and challenges associated with the TUNEL assay is essential for its effective clinical application.