J Korean Acad Conserv Dent.  2011 Nov;36(6):498-505. 10.5395/JKACD.2011.36.6.498.

Microbial profile of asymptomatic and symptomatic teeth with primary endodontic infections by pyrosequencing

• Lim SM
• Lee TK
• Kim EJ
• Park JH
• Lee Y
• Bae KS
• Kum KY

• KMID: 2176604
• DOI: http://doi.org/10.5395/JKACD.2011.36.6.498

Abstract

OBJECTIVES
The purpose of this in vivo study was to investigate the microbial diversity in symptomatic and asymptomatic canals with primary endodontic infections by using GS FLX Titanium pyrosequencing.
MATERIALS AND METHODS
Sequencing was performed on 6 teeth (symptomatic, n = 3; asymptomatic, n = 3) with primary endodontic infections. Amplicons from hypervariable region of the small-subunit ribosomal RNA gene were generated by polymerized chain reaction (PCR), and sequenced by means of the GS FLX Titanium pyrosequencing.
RESULTS
On average, 10,639 and 45,455 16S rRNA sequences for asymptomatic and symptomatic teeth were obtained, respectively. Based on Ribosomal Database Project Classifier analysis, pyrosequencing identified the 141 bacterial genera in 13 phyla. The vast majority of sequences belonged to one of the seven phyla: Actinobacteria, Bacteroidetes, Firmicutes, Fusobacteria, Proteobacteria, Spirochetes, and Synergistetes. In genus level, Pyramidobacter, Streptococcus, and Leptotrichia constituted about 50% of microbial profile in asymptomatic teeth, whereas Neisseria, Propionibacterium, and Tessaracoccus were frequently found in symptomatic teeth (69%). Grouping the sequences in operational taxonomic units (3%) yielded 450 and 1,997 species level phylotypes in asymptomatic and symptomatic teeth, respectively. The total bacteria counts were significantly higher in symptomatic teeth than that of asymptomatic teeth (p < 0.05).
CONCLUSIONS
GS FLX Titanium pyrosequencing could reveal a previously unidentified high bacterial diversity in primary endodontic infections.

Keyword

Asymptomatic; Primary apical periodontitis; Pyrosequencing; Symptomatic; Taxonomy; 16S rRNA gene

MeSH Terms

Actinobacteria
Bacteria
Bacteroidetes
Fusobacteria
Genes, rRNA
Leptotrichia
Neisseria
Polymers
Propionibacterium
Proteobacteria
Spirochaetales
Streptococcus
Titanium
Tooth
Polymers
Titanium

Figure

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  Figure 1 Plots of the numbers of different operational taxonomic units (OTUs) in infected root canals from symptomatic and asymptomatic teeth. The steepness and length of the curve indicates that a large fraction of the species diversity has not sampled. PN, asymptomatic canal; PY, symptomatic canal.

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  Figure 2 Bacterial abundance and prevalence distribution at phylum level. PN, asymptomatic canal; PY, symptomatic canal.

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  Figure 3 The 10 most abundant genera identified in asymptomatic teeth (a, PN) and the 10 most abundant genera in symptomatic teeth (b, PY) with their abundance values.

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