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

Enciso, María; Sarasa, Jonás; Agarwal, Ashok; Fernández, José Luís; Gosálvez, Jaime

doi:10.1016/s1472-6483(10)60003-x

Cited by 105 publications

(85 citation statements)

References 20 publications

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“…In mammalian sperms, DNA fragmentations can occur in 2 forms: single (SSB) and double DNA strand breaks (DSBs), and it is particularly frequent in the ejaculates of subfertile males (32).…”

Section: Sperm Dna Fragmentationmentioning

confidence: 99%

The Impact of Bacterial Infections on Human Spermatozoa

Zeyad¹,

Amor²,

Hammadeh³

2017

International Journal of Women’s Health and Reproduction Scienc

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“…In mammalian sperms, DNA fragmentations can occur in 2 forms: single (SSB) and double DNA strand breaks (DSBs), and it is particularly frequent in the ejaculates of subfertile males (32).…”

Section: Sperm Dna Fragmentationmentioning

confidence: 99%

The Impact of Bacterial Infections on Human Spermatozoa

Zeyad¹,

Amor²,

Hammadeh³

2017

International Journal of Women’s Health and Reproduction Scienc

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“…This method is called the two-tailed comet (TTC) assay and combines the alkaline and neutral versions of the original comet assay to provide information about both the amount and the type of DNA damage present in a sperm cell. 10 The type of DD present in the recovered fractions of sperm after SUP and DGC has not been examined. In the present study, we aim to examine and compare the effects of the DGC and SUP processing techniques on DNA integrity using the TTC assay and the sperm chromatin dispersion (SCD) test.…”

Section: Introductionmentioning

confidence: 99%

The ability of sperm selection techniques to remove single- or double-strand DNA damage

Enciso¹,

Iglesias²,

Galán³

et al. 2011

Asian J Androl

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A wide variety of techniques for the preparation of sperm are currently available, of which the most commonly employed are densitygradient centrifugation (DGC) and swim-up (SUP). To date, these methods appear to be effective in selecting functional sperm for assisted reproduction techniques (ART), but they may have negative effects on sperm DNA. In this study, the ability of these semen processing techniques to eliminate spermatozoa containing single-and double-strand DNA damage was assessed by the two-tailed comet assay and the sperm chromatin dispersion test in 157 semen samples from patients seeking assisted reproduction treatment. Our results indicated that SUP and DGC are equally efficient in eliminating spermatozoa containing double-strand DNA damage and sperm with highly damaged (degraded) DNA, as characterized by the presence of both single-and double-strand DNA breaks. However, DGC is more efficient than SUP in selecting spermatozoa that are free from single-strand DNA damage. Future studies should characterise the importance of the various types of DNA damage and examine the sperm processing protocols used in each laboratory to determine their ability to eliminate DNA damage and hence, prevent the potential transmission of genetic mutations via ART.

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“…This technique detects both single-stranded DNA breaks (SSB) and DSB, but would not allow for differentiation between the two. Recently a two-tailed Comet assay has been developed (Enciso et al, 2009), which by revealing the total level of both SSB and DSB in individual cells, allows for a more precise and extensive analysis of the damage. However, since the technique is based on fluorescence microscopy the slides should be analyzed by an experienced observer for interpretation of the results.…”

Section: Single-cell Gel Electrophoresis Comet Assaymentioning

confidence: 99%