Assessment of Oral Bacterial Diversity from Carious Dentin and Buccal Mucosa of Caries-Active Filipino Adults through Denaturing Gradient Gel Electrophoresis (DGGE)

Abstract

Bacteria present in the oral cavity play an important role in the progression of tooth decay or dental caries, a disease that can affect a person's work efficiency according to the Department of Health. This study aims to assess and compare the bacterial diversities in carious dentin (CD) and buccal mucosa (BM) samples of caries active Filipino Adults using PCR-DGGE. This paper is a descriptive-comparative study of bacterial profiles of two oral sites 一 carious dentin and buccal mucosa. Dentin extracts and buccal swabs were collected from three male and three female subjects; and placed in MRS broth, nutrient broth and PBS solution. CT AB DNA extraction and nested PCR of 16S rRNA gene internal hypervariable regions V3-V5 was then performed. Nested PCR products were run on an 8% denaturing gradient gel electrophoresis (DGGE) polyacrylamide gel with a 40-70% gradient. DGGE gels were analyzed using Bio-Rad Quantity One software. Dice similarity coefficients of 50.73% and 55.49% were obtained for CD and BM samples, respectively. Comparison of the two oral sites yielded a 50.26% Dice similarity coefficient. Despite the weak similarity demonstrated by the similarity coefficients, it was believed that a core group for both CD and BM samples could exist as suggested by the presence of ubiquitous and frequently recurring bands. The presence of bands ubiquitous in both sites also suggested the possible occurrence of bacterial translocation but the limited data from this study could not verify this idea. Identification of bacteria and physiological studies were needed to verify this.