TUNEL labeling with BrdUTP/anti-BrdUTP greatly underestimates the level of sperm DNA fragmentation in semen evaluation
| Autor: | Maria De Geyter, Sofia C. Ribeiro, Monica Muratori, Christian De Geyter |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Male
0301 basic medicine Physiology Condensation lcsh:Medicine Gene Expression Apoptosis DNA fragmentation Biochemistry 0302 clinical medicine Animal Cells Medicine and Health Sciences Prospective Studies lcsh:Science Staining sperm DNA fragmentation brighter and dimmer sperm population TUNEL flow cytometry 030219 obstetrics & reproductive medicine Multidisciplinary TUNEL assay Cell Death medicine.diagnostic_test Chromosome Biology Chemistry Physics Cell Staining Condensed Matter Physics Spermatozoa Chromatin Body Fluids Nucleic acids Cell Processes Physical Sciences Fluorescein Epigenetics Cellular Types Anatomy Phase Transitions Research Article DNA damage Semen Semen analysis Research and Analysis Methods Andrology 03 medical and health sciences In Situ Nick-End Labeling Genetics medicine Humans Infertility Male lcsh:R Biology and Life Sciences Cell Biology DNA Sperm Semen Analysis Germ Cells 030104 developmental biology Terminal deoxynucleotidyl transferase Specimen Preparation and Treatment lcsh:Q Deoxyuracil Nucleotides |
| Zdroj: | PLoS ONE PLoS ONE, Vol 12, Iss 8, p e0181802 (2017) |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0181802 |
| Popis: | Many studies have now confirmed that sperm DNA fragmentation (SDF) is associated with a poorer outcome of some forms of assisted reproduction technology. For this reason, SDF is an important parameter to evaluate in male fertility assessment. TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) assay coupled to flow cytometry is one of the most promising methods for SDF quantification. Several kits for the detection of DNA fragmentation are currently available on the market and all are recommended as equally appropriate to quantify SDF. In this work we compared for the first time the efficacy of two different types of TUNEL kits for SDF quantification: one using an indirect antibody-based labeling system (BrdUTP/fluorescein-anti-BrdUTP) and another using a direct labeling system (fluorescein-dUTP). We demonstrated that TUNEL indirect labeling system largely underestimates SDF when compared with the direct labeling, the differences ranging from 19.2% to 85.3% (p |
| Databáze: | OpenAIRE |
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