Quantitative analysis of Epstein-Barr virus and Periodontopathogens in Periodontitis Sites

Objectives: To investigate the levels of EB virus, P. gingivalis, T. forsythia, T. denticola, A. actinomycetemcomitans, P. intermedia and C. rectus in periodontitis sites, and to find the relationship and the role of these pathogenic agents during the progression of periodontitis. Methods: The pooled subgingival samples from the four deepest active and four disease-stable periodontitis sites for each patient of 60 chronic periodontitis patients were collected by paper points and transported into 0.9% NaCl solution. After vortexing the homogenous suspension was divided into two aliquots for the isolation of bacterial and virus DNA respectively. EB virus, P. gingivalis, T. forsythia, T. denticola, A. actinomycetemcomitans, P. intermedia and C. rectus were identified and quantified by 5'-nuclease (TaqMan) real-time PCR. The standards were established through the serial dilutions of plasmids containing the amplified region of each pathogen using cloning procedure. Results:There was no significant differences in the occurrences of each pathogen between stable and active sites (p>0.05). Further quantitative analysis showed that the infection levels of all the test species in the active periodontitis sites were significantly higher than in the stable sites (p<0.001). The significant correlations were observed between the clinical parameters mean PD, CAL and the numbers of EB virus, P. gingivalis, T. forsythia, T. denticola and C. rectus respectively. In active periodontitis sites positive correlations were found between the copy numbers of EBV and P. gingivalis, EBV and T. forsythia, EBV and C. rectus. Conclusions: The amount of microorganisms in periodontal sites might be an important determinant for the development of the disease, EBV and periodontopathic bacteria may participate in the initiation or progression of periodontits jointly. This study was supported by Grant No.7003472 of Natural Science Foundation of Guangdong Province.