Molecular Identification of Candida albicans in Endodontic Retreatment Cases by SYBR Green I Real-time Polymerase Chain Reaction and its Association with Endodontic Symptoms.

Abstract

Recent microbiological studies have expressed ever-increasing concerns about Candida albicans as a causal factor in the failure of endodontic treatments. Real-time quantitative polymerase chain reaction (qPCR), including the SYBR Green I system, is a technique in which a fluorescent dye is incorporated into the double-stranded DNA that is produced during DNA polymerase activity. This study aimed to determine the relative prevalence of Candida albicans in root canals of retreatment cases and its association with endodontic symptoms. In the present cross-sectional/analytical study fifty subjects were selected. Clinical features and radiographic status of the teeth were also evaluated. After access cavity preparation, the retrieved material and dentinal chips removed from the root canal were transferred into 1.5-mL microtubes, followed by storage at -20ºC until used for DNA extraction. A DNeasy Tissue Kit was used to extract DNA using the DNeasy protocol for animal tissues. Master Plus SYBR Green I (Jena Bioscience, Germany) was used in a Rotor-gene Real-time PCR System for real-time PCR. The relationship between the presence of Candida albicans and the clinical and radiographic features were analyzed using McNemar's test. There was a significant relationship between the radiographic findings in endodontically treated teeth and the presence of Candida albicans. However, there was no significant relationship between the presence of Candida albicans and any of the clinical symptoms. In spite of the limitations of this study, we concluded that Candida albicans was associated with root canal infections in endodontic retreatment cases, but there was no relationship between root canal infections and the clinical symptoms.