A two-tailed Comet assay for assessing DNA damage in spermatozoa

REPRODUCTIVE BIOMEDICINE ONLINE

卷:18

Article

Enciso, Maria¹  Sarasa, Jonas¹  Agarwal, Ashok²  Fernandez, Jose Luis³  Gosalvez, Jaime¹ 

[1] Univ Autonoma Madrid, Unidad Genet, Dept Biol, Madrid, Spain

[2] Cleveland Clin, Ctr Res Reprod, Cleveland, OH 44106 USA

[3] Ctr Hosp Juan Canalejo, La Coruna, Spain

关键词: DNA fragmentation;    double-stranded DNA breaks;    single cell gel electrophoresis (SCGE);    single-stranded DNA breaks;    spermatozoa;   

DOI  :  10.1016/S1472-6483(10)60003-X

来源: Elsevier

【 摘 要 】

DNA fragmentation is considered an important parameter of semen quality, and of significant value as a predictor of male fertility. Poor quality chromatin is closely associated with, and highly indicative of, some fertility problems. Many methodologies to assess DNA fragmentation in spermatozoa are available, but they are all unable to differentiate between single-stranded DNA breaks (SSB) and double-stranded DNA breaks (DSB) in the same sperm cell. The two-tailed Comet assay (2T-Comet) protocol overcomes this limitation. A modification of the original Comet assay was developed for the simultaneous evaluation of DNA SSB and DSB in human spermatozoa. The 2T-Comet assay is a fast, sensitive, and reliable procedure for the quantification and characterization of DNA damage in spermatozoa. It is an innovative method for assessing sperm DNA integrity, which has important implications for human fertility and andrological pathology.

【 授权许可】

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