Metagenome Analysis of Buccal Mucosal Biofilms to Identify Bacterial Prevalence in Subjects with and without type II Diabetes

Abstract

To quantify the two most prevalent bacteria among Type II diabetic individuals and controls from the buccal mucosal biofilms using molecular methods. To compare the percent prevalence of Veillonella and Granulicetella bacteria in uncontrolled Type II diabetic individuals with a control group. Subjects selected randomly and categorized into two groups within the age range of 25 to 40 years diagnosed with and without Type II diabetes based on their HbA1c values. The samples of buccal mucosa biofilms are collected in sterile swabs and stored in bacterial lysis buffer, which was later subjected to quantification of DNA followed by 16S rRNA amplification and sequencing. The sequence obtained is then surveyed using BLAST Analysis to define the bacterial flora and two bacteria, namely Veillonella and Granulicatella are selected for further amplification and quantification by real-time PCR to express the bacteria in copy numbers. From the collected buccal mucosal biofilm samples (n=24), which was categorized into Type II diabetes (12) and non-diabetic (12). The sequence subjected to BLAST analysis gave a List of bacteria from which Veillonella sp. and Granulicatella sp. were selected and administered to real-time PCR for amplification and quantification, which revealed an increased bacterial prevalence in Type II diabetic subjects to non-diabetic subjects which was also proved statistically. Based on the results obtained, there is a significant prevalence of bacterial content in Type II diabetic subjects compared to non-diabetic subjects.