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Journal of Clinical Microbiology, September 2001, p. 3282-3289, Vol. 39, No. 9
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.9.3282-3289.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Molecular Identification of Microorganisms from
Endodontic Infections
H. J.
Rolph,1,*
A.
Lennon,1
M. P.
Riggio,1
W. P.
Saunders,2
D.
MacKenzie,1
L.
Coldero,1 and
J.
Bagg1
Infection Research Group, University of
Glasgow Dental School, Glasgow G2 3JZ,1 and
University of Dundee Dental School, Dundee DD1
4HN,2 United Kingdom
Received 5 February 2001/Returned for modification 22 April
2001/Accepted 5 July 2001
A relatively wide range of bacteria have been isolated from root
canals using standard culture techniques. However, only 50% of the
bacteria in the oral cavity are cultivable (S. S. Socransky et
al., Arch. Oral Biol. 8:278-280, 1963); hence, bacterial diversity in
endodontic infections is underestimated. This study used a PCR-based
16S rRNA gene assay, followed by cloning and sequencing of 16S rRNA
amplicons from a small subset of samples to assess the diversity of
bacteria present in infected root canals. A total of 41 clinical
samples from 15 de novo and 26 refractory cases of endodontic
infections were assessed. Of these samples, 44% were positive by
culture and 68% were positive by PCR. Eight samples were selected for
further analysis. Of these, the two de novo cases yielded sequences
related to those of the genera Enterococcus, Lactobacillus,
Propionibacterium, and Streptococcus and two clones were related to previously uncultivated bacteria, while the
sinus-associated, de novo case yielded sequences related to those of
the genera Lactobacillus, Pantoea, Prevotella, and
Selenomonas. The five refractory cases produced clones
which were related to the genera Capnocytophaga, Cytophaga,
Dialister, Eubacterium, Fusobacterium, Gemella, Mogibacterium,
Peptostreptococcus, Prevotella, Propionibacterium, Selenomonas,
Solobacterium, Streptococcus, and Veillonella and two
clones representing previously uncultivated bacteria. The phylogenetic
positions of several clones associated with the
Clostridiaceae and Sporomusa subgroups of the
Firmicutes grouping are also shown. This study demonstrates
that molecular techniques can detect the presence of bacteria in
endodontic infections when culture techniques yield a negative result
and can be used to identify a wider range of
endodontic-infection-related bacteria including the presence of
previously unidentified or unculturable bacteria.
*
Corresponding author. Mailing address: Infection
Research Group, Level 9, Glasgow University Dental School, 378 Sauchiehall St., Glasgow G2 3JZ, Scotland. Phone: 0141 211 9742. Fax:
0141 353 1593. E-mail: H.Rolph{at}dental.gla.ac.uk.
Journal of Clinical Microbiology, September 2001, p. 3282-3289, Vol. 39, No. 9
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.9.3282-3289.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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