Reliability of bulky DNA adducts measurement by the nuclease P1 32P-post-labelling technique |
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| Authors: | R Ibáñez A Munnia A Agudo A Berenguer P Amiano M J Tormo |
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| Institution: | 1. Unit of Epidemiology, Catalan Institute of Oncology (ICO), L'Hospitalet de Llobregat, Barcelona, Spainraquelip@iconcologia.catsalut.net;3. Cancer Risk Factor Branch, Molecular Biology Laboratory, CSPO-Scientific Institute of Tuscany Region, Florence, Italy;4. Unit of Epidemiology, Catalan Institute of Oncology (ICO), L'Hospitalet de Llobregat, Barcelona, Spain;5. Dirección de Salud de Guipúzcoa, San Sebastián, Spain;6. Consejería de Sanidad y Consumo, Murcia, Spain |
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| Abstract: | The aim was to assess the reliability of bulky DNA adducts measurement by means of the 32P-post-labelling assay. The research design consisted of an intramethod reliability study. Buffy coats from 41 subjects were used to obtain two aliquots of 1–5 μg DNA for each subject; bulky DNA adducts were measured using the nuclease P1 32P-post-labelling technique. The reliability of the measurement was assessed by means of the intraclass correlation coefficient (ICC), the distribution of the differences between the two measurements and the limits of agreement. The estimated ICC was 0.977, with a 95% confidence interval between 0.921 and 0.977. The limits of agreement were ±0.44 (DNA adducts per 108 nucleotides). Only three subjects had differences lying out of such limits. Bulky DNA adduct levels measured by the 32P-post-labelling technique showed good reliability. Only one measurement is needed to use DNA adducts as a biomarker of exposure and, possibly, cancer risk. Besides, as a validation analysis, 32P-post-labelling measurements can be repeated in only 20–30% of samples. |
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| Keywords: | Reliability DNA adducts post-labelling technique |
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