DETECTION OF Aggregatibacter actinomycetemcomitans FROM PERIODONTITIS AND HEALTHY INDIVIDUALS BY CULTURE AND PCR METHODS

Objective: The aim of this study was to determine the detection of Aggregatibacter actinomycetemcomitans from periodontitis and healthy individuals by culture and PCR methods.

Methods: Clinical isolates were obtained of periodontal pockets by paper point from 50 health subjects with periodontal pockets depth < 5 mm and 50 patients with periodontitis with periodontal pockets depth equal to or exceeding 5 mm that were referred to Sadaf dental clinic in Shiraz, Iran. These samples were cultured in Tryptic Soy Agar (TSA) and incubated in an anaerobic system. A. actinomycetemcomitans were isolated and identified using star like colony structure, Gram staining and catalase test. For PCR, samples were analyzed with genus specific primers. The primers were used to multiply the region about 500 bp of A. actinomycetemcomitans.

Results: Of the 100 samples, A. actinomycetemcomitans was isolated from 33 patients (33%), (8 isolate of healthy subjects (16%); 25 isolate of patients (50%)) by using a selective Aggregatibacter isolation medium. Using PCR, a total of 49 (49%) samples were found to be positive for Aggregatibacter (12 isolate of healthy subjects (24%); 37 isolate of patients (74%)). There was a direct relationship between the depth of pocket and the presence of the A. actinomycetemcomitans (P<0.05).

Conclusions: The presence of A. actinomycetemcomitans in dental plagues from patients with periodontitis could indicate a role for periodontal pathogenic bacteria in the periodontal diseases. It is well know that improvements in diagnostic methods are useful in the prevention and treatment of periodontal diseases.