Quantitative evaluation of selected etiological factors of periodontitis in patients with atherosclerotic disease using qPCR and ddPCR methods

Project manager: dentist. Barbara Brzychczy
Implementation period: 2024-2025

The results obtained from the project (Disorders of the oral microbiome in the course of selected cardiovascular diseases, in-house research N41/DBS/000970) using next-generation sequencing (NGS) indicate the presence of marker bacteria specific to the studied groups of patients who underwent endarterectomy: I - patients with periodontal disease, II - patients without periodontal disease. Since the data are semi-quantitative and narrowed down to the genus level only (this is due to the limitation of NGS related to the much lower reliability of matching sequences to the species level), the aim of the study will be to use much more sensitive methods - ddPCR and qPCR to precisely quantify selected species representing oral pathogens obtained at the genus level. Amplification of bacterial DNA using ddPCR is a fully quantitative technology-based method, which will allow precise estimation of DNA volume at 2,000 dots per sample reaction, which, unlike NGS (which yields a number that only allows a small number of DNA copies in the sample), will allow precise assessment of the concentration of the pathogens under study in the samples. In addition, it will make it possible to obtain small numbers of DNA copy numbers without the need for reference markers. The results obtained in the study using qPCR and ddPCR will allow assessment of the degree of malignancy of both methods, and will enable analysis of the correlation between the obtained quantitative data of selected pathogenic threads with clinical indications. The results obtained on selected groups of biological materials may serve as a predictor of the clinical status of patients in both oral and atherosclerotic disease, which may serve as a prognostic marker in the future.