Methods: For the gene polymorphism studies DNA was isolated from oral mucosal scrapings collected from patients of the clinics of Semmelweis University. As a first step of our experimental series, we aimed to set and optimize combined polymerase chain reaction (PCR) and restriction fragment length polymorphism analysis (RFLP) methods to identify individual gene polymorphisms.
Results: DNA isolation was carried out after optimization of sample collection and storage protocols to prevent DNA degradation. We successfully optimized eight SNP procedures that are putatively involved in periodontitis (IL-1 alpha -889 C/T, IL-1 beta -511 C/T, IL-1 beta +3954 C/T, IL-6 -174 C/G, IL-10 -1082 G/A, TLR-4 -299 G/A, TLR-4 -399 T/C, TNF alpha -308 A/G). We observed the presence the rare allele as well as the dominant allele in each investigated putative polymorphism in the population studied, although at present we have a relatively small sample number.
Conclusion: Our results are primarily methodological and clearly show the feasibility of such studies. On this basis now we perform population studies with large sample numbers to compare polymorphisms of a large number of genes in healthy and diseased groups. Our long term goal is to identify the gene polymorphisms participating in the development of the disease by population genetics/genomics methods. These achievements may help in the primary control of periodontal diseases and also project the development of new diagnostic strategies.
Supported by the Hungarian Scientific Research Fund (OTKA 72385).