A two-tailed Comet assay for assessing DNA damage in spermatozoa.
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.
Enciso
Maria
M
Departamento de Biología, Universidad Autónoma de Madrid, Spain. mariaencisolorences@gmail.com
Sarasa
Jonas
J
Agarwal
Ashok
A
Fernández
Jose Luis
JL
Gosálvez
Jaime
J
D000165:Acridine Orange; D020552:Comet Assay / Q000379:methods*; D053938:DNA Fragmentation; D006801:Humans; D006861:Hydrogen Peroxide; D007091:Image Processing, Computer-Assisted; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D008297:Male; D012680:Sensitivity and Specificity; D013094:Spermatozoa / Q000737:chemistry*; D018709:Statistics, Nonparametric
D006861:Hydrogen Peroxide; D000165:Acridine Orange