Clinical and molecular detection of periodontopathic bacteria using universal primers in different periodontal diseases treated with non-surgical modalities

ABSTRACT

Difference in the composition of microbiota in different periodontal diseases has been observed. There are several microbial methods used to identify the periodontopathic bacteria which harbor the periodontally affected sites. The present study compared molecular detection of bacterial periodontitis to conventional microbiological method as well as identifying prominent isolated species. This study also describes clinical parameters as well as evaluating the effect of non-surgical treatment modalities [SRP] among different periodontal diseases on total viable bacterial count. Clinical parameters include probing depth [PD], clinical attachment level [CAL], bleeding on probing percent [BOP%] at baseline and three months after. Gingival crevicular fluid samples from 45 patients were collected and analyzed at day 0, 1, 30 and 90 days after SRP' treatment. The microbiologic assessment was done using universal primers for PCR detection of bacterial periodontal infection. The ability to detect bacterial infection from 45 gingival crevicular fluid samples by PCR using universal primers encoding for the 16 S rDNA was compared with bacterial culture. Bacteria were detected by PCR with 100% specificity and 77.5% sensitivity compared to culture. Detection of 16 S rDNA proved to be effective in preventing PCR inhibition and was applied to detect the occurrence of non-cultural bacteria in the tested samples. The occurrence of bacteria was decreased by non-surgical mechanical debridement [SRP] which was evaluated by total viable count technique. Frequently isolated bacteria species were identified as Porpheromonas gingivitis, Fusobacterium nucleatum, Tenner Ella forsythia, Aggregibacter actinomyctemecommitans and Prevotella intermedia. The results indicated that using universal bacterial primers is a rapid and simple method for detection of bacterial periodontitis. A favorable clinical response to these therapeutic measures can be obtained. Moreover, the bacterial count is affected significantly by non- surgical treatment modality