Molecular Diagnosis Of Some Anaerobic Bacteria From The Root Canals

Abstract

The goal of this study was for molecular diagnosis of anaerobes bacteria from the root canals. Fifty patients, ranging in age from 25 to 50, have been sent to the specialized dental clinic for root canal and periodontal pocket sampling. For anaerobic growth, the samples (paper points) were put in 10 mL of thioglycollate and vortexed for 1 minute, Plates of nutrient agar were infected using sterile spreaders with 0.1 mL of undiluted sample (10 dilution) and each of the four dilutions. Anaerobic samples were cultured for up to 48 hours on nutrient agar plates in a GasPak container. After incubation in each medium, the growth was detected. Gram staining, colony morphology observation on blood agar plates, and a biochemical identification kit were used to determine the purity of the cultures. DNA of bacteria were extracted for molecular detection of specific genes of bacteria. According to conventional techniques and biochemical analyses, 22 different strains of Enterococcus sp., 6 isolates of P. gingivalis, and 2 of P. intermediawere found in root canal samples. Traditional culture techniques and biochemical testing revealed 19 (86.3 percent) of the 22 Enterococcus spp. isolates as being of Enterococcus faecium level, whereas molecular identification identified 17 (77.3 percent) of the isolates as being of Enterococcus faecium level, while 4 from P. gingivalis and 1 from P. intermedia were confirmed by PCR. To find particular genes, all the isolates in this investigation were subjected to PCR using species-specific primers and a PCR method called PCR.

In conclusion, Enterococcus faecium was detected in high percentages as a main pathogen of root canal infection followed by P. gingivalis, and P. intermedia.