Assessment of intraradicular bacterial composition by terminal restriction fragment length polymorphism analysis

Background: The aim of the study was to assess the bacterial community structures associated with endodontic infections using terminal restriction fragment length polymorphism (T‐RFLP), and to investigate the correlation of whole community profiles with the manifestation of particular clinical feat...

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Vydané v:	Oral microbiology and immunology Ročník 24; číslo 5; s. 369 - 376
Hlavní autori:	Saito, D., Marsh, T. L., De Souza Cannavan, F., Höfling, J. F., Gonçalves, R. B.
Médium:	Journal Article
Jazyk:	English
Vydavateľské údaje:	Oxford, UK Blackwell Publishing Ltd 01.10.2009 Blackwell
Predmet:	16S ribosomal DNA Actinomyces - classification Adolescent Adult Bacteria Bacteria - classification Bacteria - genetics Bacteriology Bacteroides - classification Biological and medical sciences Campylobacter sputorum - classification Capnocytophaga - classification Community structure Dental Pulp Cavity - microbiology Dental Pulp Necrosis - microbiology Deoxyribonuclease HpaII Deoxyribonucleases, Type II Site-Specific DNA, Bacterial - genetics endodontic infection Endonuclease Eubacterium - classification Female Flavobacterium - classification Fundamental and applied biological sciences. Psychology Fusobacterium nucleatum - classification genomics Humans Infection Lactobacillus - classification Male Marshes Microbiology Middle Aged Miscellaneous Nucleotide sequence Peptostreptococcus - classification Periapical Diseases - microbiology Polymerase chain reaction Polymorphism, Restriction Fragment Length - genetics Prevotella - classification Primers Restriction fragment length polymorphism Retinoblastoma protein root canal root canals Selenomonas - classification Sequence Analysis, DNA Species richness terminal restriction fragment length polymorphism Veillonella - classification Young Adult Immunology Restriction fragment length polymorphism 16S ribosomal DNA Microbiology Root community structure Ribosomal DNA Bacteria Genotype endodontic infection root canal terminal restriction fragment length polymorphism
ISSN:	0902-0055, 1399-302X, 1399-302X
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Popis
Shrnutí:	Background: The aim of the study was to assess the bacterial community structures associated with endodontic infections using terminal restriction fragment length polymorphism (T‐RFLP), and to investigate the correlation of whole community profiles with the manifestation of particular clinical features. Methods: Intraradicular samples were collected from 34 subjects and classified into three study groups based on the observed clinical symptoms: acute (n = 16), sub‐acute (n = 8), and asymptomatic (n = 10). Genomic DNA was extracted from each sample, submitted to polymerase chain reaction using a fluorescently labeled 16S ribosomal DNA forward primer, and digested with two tetrameric endonucleases (HhaI and MspI). The terminal restriction fragments (T‐RFs) were subsequently discriminated in an automated DNA sequencer, and the results were filtered using a statistics‐based criterion. Results: Totals of 138 (HhaI) and 145 (MspI) unique T‐RFs were detected (means 13.1 and 11.9) and there was high inter‐subject variability in the bacterial assemblages. Odds‐ratio analysis unveiled the existence of higher order groups of positively associated T‐RFs, restating the concept that intricate ecological relationships may take place in the root canal space. A significantly greater T‐RF prevalence was detected in acute cases, suggesting a straight correlation between species richness and spontaneous pain. Conclusion: Overall, no T‐RFLP profile representing a specific bacterial consortium could be associated with the manifestation of symptoms of endodontic origin.
Bibliografia:	ArticleID:MOM525 istex:58EE1A4C567D42D9ED2EA627787A986C493D07F9 ark:/67375/WNG-QBP3J5D8-W ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23
ISSN:	0902-0055 1399-302X 1399-302X
DOI:	10.1111/j.1399-302X.2009.00525.x