RAPID DETECTION OF ACTINOBACILLUS-ACTINOMYCETEMCOMITANS, PREVOTELLA-INTERMEDIA AND PORPHYROMONA-GINGIVALIS BY MULTIPLEX PCR

The identification of specific periodontal pathogens by conventional m ethods, mainly anaerobic cultivation, is difficult, time consuming and even sometimes unreliable. Therefore, a multiplex PCR method for simu ltaneous detection of Actinobacillus actinomycetemcomitans (A.a.), Por phyromona gingivalis (P.g.) and Prevotella intermedia (P.i.) was devel oped for rapid and easy identification of these specific bacterial pat hogens in subgingival plaque samples. In this paper, there is a detail ed description of the oligonucleotide primer selection, DNA extraction and PCR conditions and the sequencing of the amplified products. The locus chosen to be amplified is a highly variable region in the 16S ri bosomal DNA. For the development of this technique ATCC cultures and p ure cultures from subgingival plaque samples taken from periodontitis patients were used. As an internal positive control a recombinant plas mid was developed. This simple DNA. extraction procedure and the DNA a mplification and visualization of the amplified product permits the de tection of the bacteria in a working day. Thus, this multiplex PCR met hod is a rapid and effective detection method for specific periodontal pathogens.