Genotypic analysis of saliva from DM-type-2 and periodontitis subjects

Saliva is a non-invasive and valuable source to study several diseases, including diabetes and periodontitis. Both diseases induce elevated proinflammatory cytokines and thus prolong inflammation. To date, variations in salivary components have been of interest. Here, a primer 'RAV' was originally designed from RAGE, ATP binding cassette transporter A1 and vitamin D-binding protein genes, which are involved in inflammation pathways. Objectives: The aim of the study was to determine whether there was a difference in PCR-patterns of saliva from diabetic, non-diabetic and periodontitis groups. Methods: Saliva was collected from 12 subjects, including three of each healthy, periodontitis, periodontitis-diabetic and diabetic subjects (average age 36±7 yrs). DNA was extracted using the E-zi DNA kit (Sunolin®, Thailand) and then processed to AP-PCR with the 14-oligomer ‘RAV' (annealing temperature of 50 degree celsius). After being electrophoresed on 1.5% agarose gel, PCR-patterns were densitometrically analyzed (BioRad®, Italy). Results: Seven distinct genotypes were found within the four groups. No significant correlation was observed among the patterns and clinical signs. However, a unique genotype was detected in all periodontitis-diabetic patients. Also, a few amplicons were found conserved among the diabetic and non-diabetic subjects. Conclusion: Data here implied that AP-PCR with the primer ‘RAV' to analyze genotypic traits in saliva was limited in discriminating diabetes and periodontitis. However, more samples are required to elucidate such association. This work was supported by the grant of Mahidol University, Thailand.