Determination of malondialdehyde-induced DNA damage in human tissues using an immunoslot blot assay
| Autor: | David E. G. Shuker, C. Lagneau, Chiara Leuratti, John P. Plastaras, Rajinder Singh, Lawrence J. Marnett, Peter B. Farmer |
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| Rok vydání: | 1998 |
| Předmět: |
Detection limit
Cancer Research DNA damage Immunoblotting Mutagen DNA General Medicine Malondialdehyde medicine.disease_cause High-performance liquid chromatography Molecular biology Lipid peroxidation Blot DNA Adducts Mice chemistry.chemical_compound chemistry medicine Animals Humans Chromatography High Pressure Liquid DNA Damage |
| Zdroj: | Carcinogenesis. 19:1919-1924 |
| ISSN: | 1460-2180 |
| DOI: | 10.1093/carcin/19.11.1919 |
| Popis: | Malondialdehyde (MDA) is a product of lipid peroxidation and prostaglandin biosynthesis. It is mutagenic and carcinogenic and the major adduct formed by reaction with DNA, a highly fluorescent pyrimidopurinone (M 1 -dG), has been detected in healthy human liver and leukocyte DNA. Analytical methods used so far for the detection of M 1 -dG have not been applied to a large number of individuals or variety of samples. Often, only a few μg of DNA from human tissues are available for analysis and a very sensitive assay is needed in order to detect background levels of M 1 -dG in very small amounts of DNA. In this paper, the development of an immunoslot blot (ISB) assay for the measurement of M 1 -dG in 1 μg of DNA is described. The limit of detection of the assay is 2.5 adducts per 10 8 bases. A series of human samples were analysed and levels of 5.6-9.5 (n = 8) and 3.1-64.3 (n = 42) of M 1 -dG per 10 8 normal bases were detected in white blood cell and gastric biopsy DNA, respectively. Results on four human samples were compared with those obtained using an HPLC/ 32 P-post-labelling (HPLC/PPL) method previously developed and indicated a high correlation between M 1 -dG levels measured by the two assays. The advantages of ISB over other assays including HPLC/PPL, such as the possibility of analysing 1 μg DNA/sample and the fact that it is less time-consuming and laborious, means that it can be more easily used for routine analysis of a large number of samples in biomonitoring studies. |
| Databáze: | OpenAIRE |
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