Abstract
BACKGROUND: The sirtuins are a family of highly conserved NAD+-dependent deacetylases that act as cellular sensors to detect energy availability and modulate metabolic processes. Genetic variations such as single-nucleotide polymorphisms in the SIRT1 gene have been associated with several disease-related phenotypes, including cardiovascular disease, diabetes, body mass index, obesity, cholesterol metabolism, energy expenditure, glucose tolerance. Therefore, the aim of this study was to establish a rapid and sensitive molecular based assay like ARMS-PCR system for the detection of Sirtuin 1 rs7895833A>G gene variation in the Smoker and nonsmoker population of Tabuk -Saudi Arabia.
METHODOLOGY: This study was conducted on 100 confirmed subjects of Smoker and nonsmoker population of Tabuk. DNA extraction was done by using Qiagen Kit and ARMS-PCR system was optimized to detect Sirtuin 1 rs7895833A>G gene variation in Smoker and nonsmoker population.
RESULTS: All demographic features of the subjects are depicted in table 3. This study was done on 100 subjects among whom 50 were confirmed participants of smokers and 50 were confirmed subjects of nonsmokers. Out of 50 Smokers, 15[30%] were below or equal to 40 years age and 35[70%] were above 40 years of age. Of 50 consecutive smokers, 40[80%] were males and 10 [20%] were females. Out of 50 nonsmokers or healthy controls 45[90%] were males and 05[10%] were females. Out of 50 controls, 20[40%] were below or equal to 40 years age and 30 [60%] were above 40 years of age. ARMS-PCR system was optimized to detect Sirtuin 1 rs7895833A>G gene variation in confirmed subjects of Smoker and nonsmoker population. Gradient PCR was performed for SIRT1-rs7895833A>G gene variation optimization. The technique was successfully optimized by using wild-type or mutant-type primers with matched or one-base mismatched to examine the known SNPS in SIRT-1-rs7895833A>G gene. This assay does not entail any special equipment other than a thermocycler and gel documentation system.
CONCLUSION: ARMS-PCR system for SIRT-1-rs7895833A>G gene variation was successfully optimized. The assay proved to be fast, accurate, simple and economical that does not entail any special equipment other than a thermocycler and gel documentation system. It was indicated that ARMS-PCR system can be used as a potential molecular tool for the detection of SIRT-1-rs7895833A>G gene.