Method for detecting single-strand breaks in DNA

摘要

Detection of single-strand breaks in cellular DNA by incubating a sample comprising cells containing said DNA with an alkaline medium capable of releasing DNA from the cells and at least partially unwinding said DNA, and subjecting the alkaline medium-treated sample to a sandwich immunoassay using a monoclonal antibody directed against single-stranded DNA to determine the amount of single-stranded DNA therein. The method is particularly suited for estimating the quality of sperm of vertebrates. To allow a more accurate estimation of the number of single-strand breaks in the DNA, aliquots of the sample may be subjected to a pretreatment that induces single-strand breaks, such as irradiation with γ- or X-rays, using various doses to allow making a dose-effect curve.