engleski

Polymerase chain reaction (PCR) based detection of Streptococcus mitis and Leptotrichia buccalis from periodontal pockets

Background and Purpose: Periodontal diseases are infectious diseases followed by inflammatory changes in periodontal tissues, which lead to destruction of periodontal tissue and tooth loss. Many risk factors other than bacterial infection are involved in the development and progression of the disease: e.g. stress and cigarette smoking. The aim of this study was to establish a highly specific polymerase chain reaction (PCR), assay for detection of two strains of bacteria, Streptococcus mitis and Leptotrichia buccalis, which are putative periodontal pathogens. Materials and Methods: Paper points were immersed in periodontal pockets of patients with periodontal disease in order to collect bacteria, followed by DNA extraction and PCR, which was performed with primers constructed according to published bacterial DNA sequences from the Gene Bank. The specificity of positive samples was further confirmed by cutting the amplicon with restriction endonucleases (RFLP; restriction fragment length polymorphism). results and Conclusions: the primers specific for detection of Streptococcus mitis and Leptotrichia buccalis were constructed and optimized for PCR assays. The high specificity of expected PCR amplicons was confirmed by RFLP method. Hence, we strongly recommend the PCR assay described herein for specific detection of Streptococcus mitis and Leptotrichia buccalis in periodontal pockets.

periodonttis; PCR; Streptococcus mitis; Leptotrichia buccalis

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