Arun Kashivishwanath Shettar1*, Mujeeb M. A.2, Joy H. Hoskeri1, Shivakumar B. Madagi1, Sunanda Sarwad1, Vedamurthy A. B.2
1Department of Studies in Bioinformatics and Biotechnology, Akkamahadevi Women’s University Vijayapura-586108, Karnataka, India
2Department of Biotechnology and Microbiology, Karnatak University, Dharwad-580003, Karnataka, India
*Address for Corresponding Author
Dr. Arun Kashivishwanath Shettar
Department of Studies in Bioinformatics and Biotechnology, Akkamahadevi Women’s University Vijayapura-586108, Karnataka, India
Objective: The objective of the present work was to evaluate the DNA damage in radiation workers by using single cell gel electrophoresis (comet assay). Material and Methods: A total of 30 blood samples of radiations workers from different hospitals of Dharwad and Bagalkot districts of Karnataka were collected. The blood samples of alcoholic and smoking individuals were avoided so as to emphasize only on the effect of radiation. Collected blood samples were subjected for comet assay by following standard procedure. After the comet assay, cells were stained with fluorescent dye and observed in fluorescent microscope. The DNA damage was estimated by measuring the length of the comet tail by using visual scoring method. Microscopic examination of individual leukocytes samples embedded in agarose was subjected to electrophoresis and stained with a fluorescent DNA binding dye Acridine Orange Showed smear of DNA in damaged cells based on the exposure period. Results: The duration of exposure and the kind of level of DNA damage were directly proportional, an exposure of 18 months to the radiation environment showed a visual score of 0.1 in sample S3. Contrastingly as the duration of exposure increased the level of DNA damage as assessed by the visual score also increased in sample 20 with an exposure of 102 weeks. Out of 20 samples only few samples of X-ray technicians showed the considerable damage of DNA. And other samples did not show such severe damages. Conclusion: Study concludes that high exposure to radiation is main cause for DNA damage in selected population of X-ray technicians. In the course of our experiment we came across variability in the extent of DNA damage among 20 exposed individuals which can be attributed to their occupation. Our experiment has shown a significant increase in amount of DNA damage in workers with the increase in duration of exposure.
Keywords: X-ray technicians; comet assay; Single cell gel electrophoresis; Blood lymphocyte and DNA damage