Bacterial examination of endodontic infections by clonal analysis in concert with denaturing high-performance liquid chromatography.
OBJECTIVE
The aim of this study was to examine the diversity of bacterial species in the infected root canals of teeth associated with endodontic abscesses by cloning and sequencing techniques in concert with denaturing high-performance liquid chromatography.


METHODS
Samples collected from five infected root canals were subjected to polymerase chain reaction (PCR) with universal 16S ribosomal DNA primers. Products of these PCRs were cloned and sequenced. Denaturing high-performance liquid chromatography (DHPLC) was used as a screening method to reduce the number of clones necessary for DNA sequencing.


RESULTS
All samples were positive for the presence of bacteria and a range of 7-13 different bacteria were found per root canal sample. In total, 48 different oral clones were detected among the five root canal samples. Olsenella profusa was the only species present in all samples. Porphyromonas gingivalis, Dialister pneumosintes, Dialister invisus, Lachnospiraceae oral clone, Staphylococcus aureus, Pseudoramibacter alactolyticus, Peptostreptococcus micros and Enterococcus faecalis were found in two of the five samples. The majority of the taxa were present in only one sample, for example Tannerella forsythia, Shuttleworthia satelles and Filifactor alocis. Some facultative anaerobes that are frequently isolated from endodontic infections such as E. faecalis, Streptococcus anginosus and Lactobacillus spp. were also found in this study.


CONCLUSIONS
Clonal analysis of the microflora associated with endodontic infections revealed a wide diversity of oral species.
Jacinto
R C
RC
Department of Restorative Dentistry, Endodontic Area, Piracicaba Dental School, State University of Campinas-UNICAMP, Piracicaba, SP, Brazil.
Gomes
B P F A
BP
Desai
M
M
Rajendram
D
D
Shah
H N
HN
D000038:Abscess / Q000382:microbiology*; D039903:Actinobacteria / Q000145:classification; D001406:Bacillaceae / Q000145:classification; D001419:Bacteria / Q000145:classification*; D001439:Bacteroides / Q000145:classification; D002851:Chromatography, High Pressure Liquid; D002999:Clone Cells; D017931:DNA Primers; D004275:DNA, Ribosomal; D003788:Dental Pulp Diseases / Q000382:microbiology*; D013293:Enterococcus faecalis / Q000145:classification; D005051:Eubacterium / Q000145:classification; D005673:Fusobacterium / Q000145:classification; D016965:Gram-Negative Anaerobic Straight, Curved, and Helical Rods / Q000145:classification; D006094:Gram-Positive Bacteria / Q000145:classification; D006801:Humans; D007778:Lactobacillus / Q000145:classification; D009691:Nucleic Acid Denaturation; D010462:Peptostreptococcus / Q000145:classification; D016133:Polymerase Chain Reaction; D016966:Porphyromonas gingivalis / Q000145:classification; D012336:RNA, Ribosomal, 16S; D013211:Staphylococcus aureus / Q000145:classification; D034366:Streptococcus anginosus / Q000145:classification
D017931:DNA Primers; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S