Endodontology
Catonella morbi and Granulicatella adiacens: new species in endodontic infections

https://doi.org/10.1016/j.tripleo.2005.09.021Get rights and content

Objective

This study intended to examine samples from primary endodontic infections for the presence of Catonella morbi and Granulicatella adiacens, 2 species that have been recently suggested to be involved with infections in other oral sites.

Study design

Genomic DNA was isolated directly from samples taken from teeth with different forms of apical periodontitis, and a devised culture-independent 16S rRNA gene-based heminested PCR assay was used to determine the prevalence of these 2 target species. Species-specific primers were developed by aligning closely related 16S rRNA gene sequences. Species-specificity for each primer pair was confirmed by running PCR against a panel of oral bacteria and by sequencing of DNA from representative positive samples.

Results

C morbi and G adiacens were detected in 33% and 19%, respectively, of the root canals associated with chronic apical periodontitis; 30% and 10%, respectively, of the cases diagnosed as acute apical periodontitis, and 16% and 11%, respectively, of the pus samples taken from acute apical abscesses. Overall, C morbi occurred in 26% and G adiacens in 14% of the samples taken from primary endodontic infections.

Conclusions

Our findings demonstrate that C morbi and G adiacens can take part in the microbiota associated with primary endodontic infections, and their specific role in the disease process warrants further elucidation.

Section snippets

Material and methods

Samples collected for previous investigations13, 14 were stored and available for reanalysis in this study. Only single-root teeth from adult patients (ages ranging from 18-60 years), all of them having carious lesions, necrotic pulps, and radiographic evidence of periradicular disease, were included in this study. Selected teeth showed an absence of periodontal pockets deeper than 4 mm. In general, 50 samples of infections of endodontic origin were obtained. According to the forms of

Results

All sample extracts were positive after the first round of PCR amplification by using broad-range 16S rRNA gene primers, indicating that the DNA extraction procedure was adequate, that bacteria were present in all examined samples, and that inhibitors of the PCR reaction were not present. No PCR products were observed in negative controls that used sterile ultrapure water instead of sample.

The specificity of each primer was tested against a panel of representative oral bacteria. The use of each

Discussion

The present study used a 16S rRNA gene-based heminested PCR protocol to detect C morbi and G adiacens in samples taken from endodontic infections associated with different forms of apical periodontitis. Utilization of a nested PCR protocol is justified by the increased sensitivity and specificity of the assay when compared to single PCR. Increased sensitivity is a function of the large total number of cycles used in nested PCR assays. In addition, target DNA is amplified in the first round of

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    This study was supported by grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), a Brazilian governmental institution.

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