PCR reaction and dot-blot hybridization to monitor the distribution of oral pathogens within plaque samples of periodontally healthy individuals

Abstract

The purpose of this study was to determine the distribution of the putative periodontal pathogens Prevotella intermedia, Prevotella nigrescens, the three oral Capnocytophaga species (C. ochracea, C. sputigena, C. gingivalis), as well as Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans in plaque samples of periodontally healthy individuals. We chose a newly developed 16S rDNA directed polymerase chain reaction (PCR) and a previously described dot-blot hybridization assay to detect, differentiate, and quantify these bacteria directly in clinical samples. The subjects of these investigations were 66 sulcus fluid samples from 17 children (ages 3 to 5) attending a kindergarten, 48 sulcus fluid samples from 12 children (ages 9 and 10) from a primary school, and 25 subgingival plaque samples isolated from 6 different periodontally healthy dental students (ages 24 to 27). We were able to demonstrate the presence of P. nigrescens in 54 (kindergarten: 5; primary school: 33; students: 16) samples by PCR and quantified it by dot-blot hybridization. In addition, we found C. ochracea in 12 (kindergarten: 2; primary school: 10) samples by PCR reaction only. The other tested bacterial species were absent by the methods used. Furthermore we confirmed the specificity of our P. nigrescens-PCR in selected samples by enzyme electrophoresis.