Clonality of Fusobacterium nucleatum in root canal infections

Abstract

Fusobacterium nucleatum is a gram-negative non-spore-forming, non-motile, obligate anaerobic rod that is normally isolated from the oral cavity. Several studies have reported a significant heterogeneity within the F. nucleatum species. The aim of the present study was to analyze the clonal diversity of F. nucleatum strains isolated from intracanal infections and to evaluate the presence of Enterobacterial Repetitive Intergenic Consensus (ERIC)-like sequences in the genome of F. nucleatum. Samples were collected from 13 single-root teeth from adult patients, all having carious lesions, necrotic pulps and radiographic evidence of periradicular bone loss. F. nucleatum was isolated from two different patients (subjects 5 and 7) by culture. Amplification of 19 colonies from subject 5 and 15 colonies from subject 7 using ERIC primers resulted in four clonal types, two per subject. An intense amplicon of approximately 700 bp was generated by ERIC-PCR for all F. nucleatum isolates and F. nucleatum ssp. polymorphum ATCC 10953. The amplification reaction using primer 1254 confirmed the results obtained with the ERIC primer. Our findings indicate that DNA fingerprints provided by ERIC- and Arbitrarily Primed (AP)-PCR may constitute a powerful tool for investigating F. nucleatum clonal diversity.

Prevalence and Clonal Analysis of Porphyromonas gingivalis in Primary Endodontic Infections

“…This is very similar to a previous study examining the clonal diversity of Fusobacterium nucleatum in 2 infected canals, in which 4 clonal types, 2 per individual, were disclosed by ERIC and AP-PCR analyses (33). Here, no P. gingivalis clonal type was shared by the 2 samples from nonrelated individuals, confirming the broad genetic diversity of the species.…”

Clonal Persistence of Oral Fusobacterium nucleatum in Infancy

“…The stability of AP-PCR types increased after the first year of life. AP-PCR typing is a simple, rapid, and reliable method for fingerprinting bacterial strains; however, its usage for F. nucleatum has thus far been limited (George et al, 1997;Avila-Campos et al, 1999;Moraes et al, 2002). We selected 4 out of 12 tested primers for AP-PCR typing of clinical F. nucleatum isolates, based on their discriminating AP-PCR patterns for different Fusobacterium strains (George et al, 1997;Narongwanichgarn et al, 2001).…”

“…In addition, some strains among F. nucleatum populations are capable of producing ␤lactamases (Könönen et al, 1999a;Nyfors et al, 2003). Using AP-PCR and enterobacterial repetitive intergenic consensus PCR to type F. nucleatum from root canal infections in two patients, Moraes et al (2002) found a single clone to be highly dominant in these infections, suggesting a difference in virulence for different clones of F. nucleatum. In children, F.…”

Endodonti̇k Enfeksi̇yonlarda Mi̇krobi̇oloji̇k Geneti̇k Tani Yöntemleri̇ni̇n Rolü

“…39,40 Endodontik mikroflorada sıklıkla bulunan Fusobacterium nucleatum'un aynı enfekte kanalda farklı klonal tiplerinin izole edilebileceği PCR esaslı moleküler yöntemler aracılığıyla yapılabilmiştir. 41 Uzun yıllar boyunca spiroketler, endodontik enfeksiyonlardan alınan örneklerde mikroskop kulanılarak gözlemlenebilmiş olsalar da tür seviyesinde tanımlanmaları yapılamamıştır. 42,43 Yapılan farklı moleküler çalışmalar endodontik enfeksiyonlarda bulunan spiroketlerin treponema türüne ait olduğunu göstermiştir.…”