Analyzing Endodontic Infections by Deep Coverage Pyrosequencing

Bacterial diversity in endodontic infections has not been sufficiently studied. The use of modern pyrosequencing technology should allow for more comprehensive analysis than traditional Sanger sequencing. This study investigated bacterial diversity in endodontic infections through taxonomic classifi...

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Bibliographic Details
Published in:Journal of dental research 2010-09, Vol.89 (9), p.980-984
Main Authors: Li, L., Hsiao, W.W.L., Nandakumar, R., Barbuto, S.M., Mongodin, E.F., Paster, B.J., Fraser-Liggett, C.M., Fouad, A.F.
Format: Article
Language:English
Subjects:
Bacterial Typing Techniques
Bacteroidetes - isolation & purification
Biological and medical sciences
Dental Pulp Diseases - microbiology
Dentistry
DNA, Bacterial - analysis
Facial bones, jaws, teeth, parodontium: diseases, semeiology
Humans
Medical sciences
Non tumoral diseases
Otorhinolaryngology. Stomatology
Research Reports
RNA, Ribosomal, 16S - genetics
Sequence Analysis, DNA - methods
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Summary:Bacterial diversity in endodontic infections has not been sufficiently studied. The use of modern pyrosequencing technology should allow for more comprehensive analysis than traditional Sanger sequencing. This study investigated bacterial diversity in endodontic infections through taxonomic classification based on 16S rRNA gene sequences generated by 454 GS-FLX pyrosequencing and conventional Sanger capillary sequencing technologies. Sequencings were performed on 7 specimens from endodontic infections. On average, 47 vs. 28,590 sequences were obtained per sample for Sanger sequencing vs. pyrosequencing, representing a 600-fold difference in “depth-of-coverage”. Based on Ribosomal Database Project (RDP II) Classifier analysis, pyrosequencing identified 179 bacterial genera in 13 phyla, which was significantly more than Sanger sequencing. The phylum Bacteroidetes was the most prevalent bacterial phylum. These results indicate that bacterial communities in endodontic infections are more diverse than previously demonstrated. In addition, deep-coverage pyrosequencing of the 16S rRNA gene revealed low-abundance micro-organisms with potential clinical implications.
ISSN:0022-0345
1544-0591
DOI:10.1177/0022034510370026