Detection of some pathogen bacteria which causes periapical diseases at human teeth by polymerase chain reaction and quantitative detection of Porphyromonas gingivalis by real time polymerase chain reaction

ABSTRACTAcute apical inflammation which is a localized or defused liquefaction lesion has caused periapical tissue damage. Although it has been claimed that infections has become due to the bacteria and nonbacterial irritants as well dental pulp and periapical tissue infections, microorganisms has been accepted as the most important etiologic factors at these kind of infections. The presence of bacteria in the canal may cause periapical lesion, too. For this reason the bacteria must be removed from the interdentinal or the root canal system. So, the treatment will be successful. There are many bacteria existing in the root canal that may lead to inflammation (abscess) and this flora contains either aerobic or anaerobic microorganisms. It has been reported that anaerobic microorganisms were mostly responsible for the root canal infections and periapical diaesases. Recently, because of the inappropriate usage of the antibiotic and the elevation in the number of multidrug resistant microrganisms in our society, there may be failures in the treatment if the aerobic and anaaerobic bacteria couldn?t be removed of the root canal. Especially for the resistant and recurrent infections, type identifications of the microorgnisms in the root canal shall be important for the antibiotic selection in treatment. Thus, classical, conventional microbiological methods such as culture are needed. But this will take time because of the fastitious and/or slowly growing bacteria and also some of them are uncultivable bacteria. Besides for identification of the species within a mix population/flora will be needed more media, chemical agents and identification kits. So this will take long time and cost will be higher. On the other hand, it was impossible to type every isolated strains by in vitro cultures. Polimerase chain reaction (PCR) is accepted as a rapid, specific, sensitive and reliable technique which enables identification of the species even among a mixed flora specifically. Real time PCR (qPCR) technique is a kind of PCR technique which gives a quantitative results besides typing the microrganisms. These techniques will be used in this study. Interdentinal samples taken from the patients aseptically from the root canal of the patients who has pulpal and periapical infections in Ankara University Faculty of Dentistry, Department of Endodontics. Then the bacterial DNA were isolated from the samples and amplification of the DNA samples with oligonucleotid primers targeted to the type specific region of 16S rRNA of Porphyromonas gingivalis strain in the study. Then electrophoresis of the PCR samples in agarose gel and imaging of them were done. The PCR products will be processed with the real time PCR technique by using species-specific oligonucletide primers with a dye in 5?-3? in the lightcyler 480 quantitatively (Roche®, İsviçre).Key Words: Molecular Cell Biology, Endodontic Microorganisms, Real Time PCR(qPCR)