Eureka! Institutional Repository

This diploma thesis focused on finding the Helicobacter pylori Hsp60 gene. This gene encodes the Heat-Shock Protein 60 protein (HSP60), which generally prevents the destruction of heat-resistant proteins [20], displays gastroduodenal diseases, autoimmune disorders [26] but more specifically, HSP60 activates NF-kappaB thus causing interleukin-8 production to increase through the Toll-like receptors of gastric epithelial cells, with the consequence of gastric inflammation [2]. There is also a possibility that it is somehow related to the attachment of H. pylori to human gastric epithelial cells [25]. 57 samples of oral wash were collected from a random sample of a general population aged 19-64, in which isolation of the bacterial DNA from two different kits was performed, first using the "Genomic DNA Purification Kit" and then using the "ExtractMe Genomic DNA Kit" reagent to verify the DNA isolation capability of the samples mouthwash. After the quantitative and qualitative determination of the bacterial DNA by the spectrophotometric method, the nested-PCR method was then applied to target the Hsp60 gene and finally, horizontal electrophoresis was performed on a 1.5% w/v agarose gel. It was found that using the "Genomic DNA Purification Kit", 5 out of 57 samples were found positive for H. pylori infection, corresponding to 8.7%, while using the "ExtractMe Genomic DNA Kit" 13 out of 57 samples were found positive, corresponding to 22.8%. In conclusion, the percentages are relatively low, which may be due to the low number of samples, the good hygiene of the teeth participants, the multiple refrigeration-thaw cycles of the samples as used for other investigations, and the selection of the right DNA isolation kit that has greater sensitivity. The interesting presentation will expand our study with more samples to draw safer conclusions on both the reliability of detection methods and the rates of bacterial origin in the cavity mouth of people of general population.