Dialister pneumosintes and Filifactor alocis have been recently considered as candidate endodontic pathogens. In this study, we
devised a 16S rDNA-directed multiplex PCR protocol for simultaneous detection of these
two bacterial species in endodontic infections. Samples were taken from infected root
canals associated with asymptomatic periradicular lesions as well as from cases of
acute periradicular abscesses. DNA extracted from the samples was used as template
for simultaneous detection of D. pneumosintes and F. alocis through a multiplex PCR assay. Two fragments of the expected sizes, one specific
for D. pneumosintes and the other for F. alocis, were simultaneously amplified from a mixture of reference genomic DNA containing
DNA from both species. Clinical samples that were positive for the target species
showed a single band of the predicted size for each species. D. pneumosintes was detected by multiplex PCR in 11 samples (7 asymptomatic and 4 abscesses) and
F. alocis was identified in 9 cases (6 asymptomatic and 3 abscesses). Six samples (3 asymptomatic
and 3 abscesses) shared the two species. Data from the present study confirmed that
D. pneumosintes and F. alocis are common members of the microbiota present in primary endodontic infections and
thereby may participate in the pathogenesis of periradicular lesions. The proposed
multiplex PCR assay is a simple, rapid, and accurate method for the simultaneous detection
of these two candidate endodontic pathogens.
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Article info
Footnotes
Supported in part by grants from CNPq, a Brazilian Governmental Institution.
Address requests for reprints to Dr. José F. Siqueira Jr., R. Herotides de Oliveira 61/601, Icaraí, Niterói, RJ, Brazil 24230-230. e-mail: [email protected]
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© 2004 The American Association of Endodontists. Published by Elsevier Inc. All rights reserved.
