Analysis of Microflora Associated With Endodontic Infection Using PCR-DGGE Profiling
Objective:The purpose of this study was to utilize Polymerase Chain Reaction and Denaturing Gradient Gel Electrophoresis (PCR-DGGE) fingerprinting, a culture-independent high throughput molecular technique, to identify the microbial profile in the infected root canals. This allows comparison of the diversity of the bacterial profiles in the different parts of the root canal with a special focus on the apical portion of the root canal in primary and secondary infections. Methods:Extracted teeth with periapical radiolucency (apical periodontitis) were sectioned and microbial samples were collected from the following segments: carious dentin,the coronal, middle, and apical portions of infected root canals of primary and secondary endodontic infections. Teeth without periradicular pathology served as controls. DNA was extracted and subjected to PCR amplification using universal bacterial 16S ribosomal RNA primers. The PCR products were fractionated by PCR-DGGE, and the electrophoretic profiles were compared among the samples. Isolated PCR products served to identify the corresponding bacterial species by comparison with different Databases. Results:Samples from the different segments displayed diverse bacterial profiles in the individual root canal system. The Microbial profile in primary endodontic infection is more diverse than those involved in failed root canal treatment cases. Profile comparison revealed that no distinct bacterial composition but different polymicrobial clusters are present in diseased root canals. The suspected endodontic pathogen Enterococcus faecalis was not found to be a prominent member of the polymicrobial communities associated with endodontic infections. Conclusion:PCR-DGGE analysis allows for high throughput screening of bacterial profiles associated with endodontic infections. No distinct single species but a variety of polymicrobial clusters are present in diseased sections and primary infection generally harbor a more diverse bacterial community compared to secondary infections.
Division: IADR/AADR/CADR General Session
Meeting:2011 IADR/AADR/CADR General Session (San Diego, California) Location: San Diego, California
Year: 2011 Final Presentation ID:1372 Abstract Category|Abstract Category(s):Microbiology / Immunology
Authors
Wang, Renke
( University of California - Los Angeles, Los Angeles, CA, USA
)
Young, Sung-ming
( University of California - Los Angeles, Los Angeles, CA, USA
)
Wang, Jen-kuei
( University of California - Los Angeles, Los Angeles, CA, USA
)
He, Xuesong
( University of California - Los Angeles, Los Angeles, CA, USA
)
Lux, Renate
( University of California - Los Angeles, Los Angeles, CA, USA
)
Chugal, Nadia
( University of California - Los Angeles, Los Angeles, CA, USA
)
Shi, Wenyuan
( University of California - Los Angeles, Los Angeles, CA, USA
)