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Human sperm DNA integrity assessed by the Comet and ELISA assays

Biomedical Sciences Research Institute Computer Science Research Institute Environmental Sciences Research Institute Nanotechnology & Advanced Materials Research Institute

Hughes, CM, McKelvey-Martin, Valerie and Lewis, SEM (1999) Human sperm DNA integrity assessed by the Comet and ELISA assays. MUTAGENESIS, 14 (1). pp. 71-75. [Journal article]

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Abstract

DNA integrity in sperm is essential for the accurate transmission of genetic information and therefore the maintenance of good health in future generations. The ELISA and Comet assays, two techniques that detect DNA damage in cells, are compared in this study of DNA integrity in human sperm. Both techniques rely on alkaline unwinding for the release of single strands of DNA from the nucleus. The ELISA detects single strands immunochemically whereas the Comet assay measures single strands drawn out by electrophoresis, stained with ethidium bromide and quantified by image analysis. The two techniques, both modified for use with sperm, detect similar levels of baseline DNA damage along with similar dose-dependent patterns of induced damage by X-ray irradiation at 10 and 30 Gy (P < 0.05). The assays are also comparable in the detection of a significant protective effect by ascorbic acid (300 and 600 mu M) and alpha-tocopherol (30 and 60 mu M) on DNA integrity, both at baseline levels and following X-ray irradiation (p < 0.01). The advantages and disadvantages of each technique are discussed.

Item Type:Journal article
Faculties and Schools:Faculty of Life and Health Sciences
Faculty of Life and Health Sciences > School of Health Sciences
Faculty of Life and Health Sciences > School of Pharmacy and Pharmaceutical Science
Research Institutes and Groups:Biomedical Sciences Research Institute
Biomedical Sciences Research Institute > Pharmaceutical Science and Practice
Institute of Nursing and Health Research > Centre for Health and Rehabilitation Technologies
ID Code:9260
Deposited By:Professor Valerie McKelvey-Martin
Deposited On:22 Jan 2010 12:28
Last Modified:04 Dec 2012 12:27

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