Detection of Actinobacillus actinomycetemcomitans from periodontitis and healthy individuals by culture and PCR methods

Abstract

The aim of this study was to determine the detection of Actinobacillus actinomycetemcomitans from periodontitis and healthy individuals by culture and PCR 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 Dental clinic of Imam Khomeini. These samples were cultured in the culture media 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. Of the 100 samples, A. actinomycetemcomitanswas isolated from 31 patients (31%), (7 isolate of healthy subjects (14%); 24 isolate of patients (48%)) by using a selective actinobacillus isolation media. Using PCR, a total of 49 (49%) samples were found to be positive foractinobacillus (14 isolate of healthy subjects (28%); 35 isolate of patients (70%)).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. Key word: Periodontitis, Actinobacillus actinomycetemcomitans, Culture, PCR.