Searching of Standard Comet Assay Parameters for Detecting Lymphocyte DNA Damages Using Fourteen Different Test Conditions
Journal Title: JOURNAL OF ADVANCES IN BIOLOGY - Year 2013, Vol 3, Issue 3
Abstract
The alkaline single cell gel electrophoresis technique (SCGE), also known as the comet assay, is an evaluation method for estimating DNA damage and genotoxicity. This assay is used to measure DNA single-strand breaks, alkali-labile sites and DNA cross-linking in individual cells. Although the comet assay is a simple, sensitive, and rapid method to compare the results of previous studies, it is often difficult to interpret because of the variability among protocols. Standardisation and validation of the comet assay could provide invaluable information for the identification of hazardous substances and the risk assessment of environmental and occupational exposure. Further use of a standardised comet assay could include monitoring the effectiveness of medical treatment and the investigation of individual variation in response to DNA damage that may reflect genetic or environmental influences. In this study, the comet assay was evaluated for 4 parameters, including the tail moment (TM), tail intensity (TI), olive tail moment (OTM) and DNA tail (DNAt) length. 6 blood samples were collected from genetically unrelated healthy volunteers, and 7 different blood volume ranges (2.5 ml to 2.5 µl, diluted with PBS or RPMI) were tested for these parameters. The intraclass correlation coefficients (ICCs) were statistically significant among all 14 different test conditions (r=0.87, p<0.001 for TM; r=0.86, p<0.001 for TI; r=0.73, p<0.05 for OTM and r=0.87, p<0.001 for DNAt). In conclusion, the determination of DNA damage using the comet assay with small blood sample volumes can contribute worthwhile knowledge to future studies.
Authors and Affiliations
Zeliha Kayaalti, Esma Soylemez, Serap Yalcin, Tulin Soylemezoglu
Keywords
Related Articles
- EP ID EP654354
- DOI 10.24297/jab.v3i3.1928
- Views 212
- Downloads 0
Surveillance ecological study of cellular responsesin three marine edible bivalve species to Cd present in their marine habitat,Mediterranean sea in Alexandria, Egypt.
Coastal habitats experience large fluctuations of environmental factors such as salinity; they also serve as the natural sinks for trace metals. Bivalves are exposed to the elevated concentrations of metals in water that...
Characterization of Phospholipase A2 Enzyme Activity in Serum of the Komodo Dragon (Varanus Komodoensis)
Soluble phospholipase A2 (sPLA2) is an enzyme found in the peripheral circulation of vertebrates which has significant immunological activity. This enzyme exerts immune activity by the hydrolysis of fatty acids fro...
UNUSUAL ASPHYXIAL DEATHS - Case Reports
Violent mechanical asphyxiation as a cause of death in the forensic practice is frequent and usual occurrence, notwithstanding whether it is a homicide, suicide or an accident. Purpose of this paper is to present unusual...
Attempt to understand the relationship between the water of the soil system and the vegetation: the case of the Martinique lower vegetation floor
The soil and water dynamics and the dynamics between soil and vegetation are difficult to explore. Yet plant phenology is closely linked to this interdependence whose characteristics vary with the seasons and the climate...
Comparison of Real time PCR with traditional tools for diagnosis of Trichomonas vaginalis in patient with vaginal discharge at Kut city, Iraq
Trichomoniasis is a sexually transmitted disease (STD) and most common form of STD which is caused by Trichomonas vaginalis. This is a descriptive analytical study. Vaginal swabs were collected from sixty female patients...