engleski

The single cell gel elecrophoresis (comet assay) for detection of DNA damage in human genome

The Comet assay or single cell gel (SCG) assay is a new, rapid and sensitive technique for visualising and measuring DNA damage and repair in individual cells. This technique detects DNA strand breaks, alkali labile damage, incomplete excision repair sites and DNA:DNA crosslinking at the level of individual cells. Its use has increased significantly in the past few years. This technique was used to study peripheral blood cells from control subjects and subjects employed in application of antineoplastic drugs. Cells embedded in agarose on microscope slides were subjected to lysis, unwinding of DNA and electrophoresis at high pH. Comets were analysed using a fluorescence microscope and Image analysis system Comet Assay II. Our results clearly indicate that lymphocytes from exposed subjects have a greater migration of DNA (measured with the comet assay as tail length, tail moment and tail % DNA) then control lymphocytes. These preliminary results showed that antineoplastic drugs caused damages of DNA which could be easily detected with comet assay. In conclusion, the comet assay might be helpful in analysing chemical and physical characteristics of genotoxic agents and applied as a standard biomonitoring tool.

genome damage; comet assay; human lymphocyte; antineoplastic drugs

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