Selected Endodontic Pathogens in the Apical Third of Infected Root Canals: A Molecular Investigation

Siqueira, Jr . José F.; Rôças, Isabela N.; Alves, Flávio R.F.; Santos, Kátia Regina Netto dos

doi:10.1097/01.don.0000125875.88377.85

Cited by 65 publications

(69 citation statements)

References 28 publications

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“…45,2007 MICROBIOTA OF ENDODONTIC INFECTIONS 3045 on May 9, 2018 by guest http://jcm.asm.org/ commonly been isolated from root canal infections (4,30,45,58). In this study, F. nucleatum subspecies were present in relatively high proportions and levels.…”

Section: Discussionmentioning

confidence: 58%

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Use of Multiple-Displacement Amplification and Checkerboard DNA-DNA Hybridization To Examine the Microbiota of Endodontic Infections

Brito

Teles

Teles³

et al. 2007

J Clin Microbiol

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Multiple-displacement amplification (MDA) has been used to uniformly amplify bacterial genomes present in small samples, providing abundant targets for molecular analysis. The purpose of this investigation was to combine MDA and checkerboard DNA-DNA hybridization to examine the microbiota of endodontic infections. Sixty-six samples were collected from teeth with endodontic infections. Nonamplified and amplified samples were analyzed by checkerboard DNA-DNA hybridization for levels and proportions of 77 bacterial taxa. Counts, percentages of DNA probe counts, and percentages of teeth colonized for each species in amplified and nonamplified samples were computed. Significance of differences for each species between amplified and nonamplified samples was sought with Wilcoxon signed-rank test and adjusted for multiple comparisons. The amount of DNA in the samples ranged from 6.80 (؎ 5.2) ng before to 6.26 (؎ 1.73) g after MDA. Seventy of the 77 DNA probes hybridized with one or more of the nonamplified samples. All probes hybridized with at least one sample after amplification. Most commonly detected species at levels of >10 4 in both amplified and nonamplified samples were Prevotella tannerae and Acinetobacter baumannii at frequencies between 89 and 100% of samples. The mean number of species at counts of >10 4 in amplified samples was 51.2 ؎ 2.2 and in nonamplified samples was 14.5 ؎ 1.7. The endodontic microbiota was far more complex than previously shown, although microbial profiles at teeth with or without periradicular lesions did not differ significantly. Species commonly detected in endodontic samples included P. tannerae, Prevotella oris, and A. baumannii.The microbiology of endodontic infections has been studied for many years (4, 47, 58). However, the association between specific microorganisms found in root canals and the symptoms of endodontic infections is poorly understood. Early studies of the endodontic microbiota indicated a predominance of aerobic and facultative bacterial species (16). This conclusion was questioned by the development of anaerobic culturing techniques which clarified the etiopathogenesis of endodontic infections by demonstrating the common occurrence of obligate anaerobic bacteria (4, 23, 30). Nevertheless, culture-based techniques have limitations, such as the difficulty in detecting fastidious anaerobic microorganisms and moderate sensitivity and specificity (44).Recently, molecular biology techniques have provided a more cost-effective, specific, and sensitive method to evaluate the microbiological profiles of oral pathologies, including endodontic and periodontal infections (37,38,44,(52)(53)(54). This technology permits the detection of microbial species that are difficult to grow as well as uncultivated and unrecognized phylotypes (34), which would lead to a better understanding of the oral microbiota, including endodontic infections (19,35,49,56).Checkerboard DNA-DNA hybridization is a high-throughput method to analyze large numbers of DNA samples by use of a wide range of...

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Section: Discussionmentioning

confidence: 58%

“…Since checkerboard DNA-DNA hybridization can detect 10 4 bacterial cells, it is possible that 10 cells of a species were detected. PCR can also detect few cells (18,19,45) but requires specific primers and may lead to major amplification bias (11,60), which is much lower in MDA (60).…”

Section: Discussionmentioning

confidence: 99%

Use of Multiple-Displacement Amplification and Checkerboard DNA-DNA Hybridization To Examine the Microbiota of Endodontic Infections

Brito

Teles

Teles³

et al. 2007

J Clin Microbiol

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“…Tannerella.forsythia'nın enfekte kök kanallarında PCR kullanılarak saptanmış prevalansları, çalışmalarda % 4-66 arasında dağılım göstermektedir (1,(5)(6)(7)(8)(9)(10)(11). Bu çalışmada saptanan prevalans % 31 olup aynı PCR tekniği ve örnekleme prosedürü kullanan diğer çalışma-ların bulgularıyla uyum göstermektedir (1,6).…”

Section: Discussionunclassified

“…PCR ile Tannerella forsythia, Treponema denticola, Dialister pneumosintes and Prevotella tannerea gibi mikroorganizmaların kök kanallarında, kültür yön-temleri ile saptanandan çok daha yüksek prevalanslarda bulundukları ortaya çıkmıştır (5,6). PCR kullanılarak yapılan farklı çalışma-larda T.forsythia'nın prevalansı %4-52 arasında değişmektedir (1,(5)(6)(7)(8)(9)(10)(11). Ayrıca bu bakterinin varlığı ile belirli semptomlar arasında bir ilişki-nin bulunup bulunmadığı henüz bir netlik kazanmamıştır.…”

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Primer kok kanal enfeksiyonlarında Tannerella forsythia prevalansının 16S rRNA PCR ile incelenmesi

ULUSOY¹,

Özgür²,

ENGİN³

et al. 2010

Ankara Üniversitesi Diş Hekimliği Fakültesi Dergisi

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The purpose of this study was to assess the prevalence of Tannerella forsythia in the primary infections of root canals and its association with different forms of periradicular diseases using Polymerase Chain Reaction (PCR).Samples were collected from acute and chronic forms of periradicular diseases and abscesses. DNA extraction was performed using spin column DNA isolation kit and 16S rRNA gene based PCR analysis was performed. Chi-squared test was used for data analysis.Tannerella forsythia DNA was detected in 18 (31,3%) of the total 58 samples, in 13 of 26 cases (50%) with acute apical periodontitis, three of seven cases (42,8%) with acute periradicular abscesses, and 2 of 25 cases (8%) with chronic asymptomatic periradicular lesions. It was found to be significantly related with acute apical periodontitis group (p=0.002).T. forsythia must be considered as a potential endodontic pathogen associated with acute symptoms of periradicular disease.

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“…Cryotubes containing the TE buffers were used for the subsequent incubation of the files and paper pints in the incubation cabinet. Subsequent deoxyribonucleic acid (DNA) extraction from the samples was done following the procedure described by Siqueira et al 10 For making the control group, several bacterial DNA was chosen as previously described in the literature. 11 Paster et al's modification of the reverse-capture checkerboard assay was used in the present study.…”

Section: Sample Collection and Dna Analysismentioning

confidence: 99%

Evaluation of Bacteriological Profile in the Apical Root Segment of the Patients with Primary Apical Periodontitis

Tatikonda

Sudheep

Biswas

et al. 2017

The Journal of Contemporary Dental Practice

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Introduction: Apical periodontitis usually results from bacterial accumulation and contamination occurring in the root-canal system, and extending beyond the apical foramen to involve the periapical tissues. Literature has a paucity of the studies that stress on the division and analysis of the pulp canal segments. The reason for this disparity might be the technique used for collecting the samples from the pulp canals. Hence, we carried out the present study to evaluate the microbial flora in the apical part of the roots with necrotic pulp canals. Materials and methods:The present study included the assessment of 40 freshly extracted teeth that had necrotized pulpal tissue along with the presence of periapical periodontal lesions. Removal of the soft tissue lesions attached to the root portion of the teeth along with apical periodontal lesions was done with the help of scalpel blade, after rinsing them with a sterile solution of saline. Thorough cleaning of the root surfaces was done with hydrogen peroxide followed by rapid disinfection with the help of sodium hypochlorite at varying concentrations. Sectioning of the root portion of all the specimens with the help of a disk was done perpendicular to the long axis of the teeth at a distance of roughly 5 to 6 mm from the teeth's apicalmost point. Cryotubes were used for transferring the specimens of apical portions containing 1 mL of buffer and were subjected to immediate frozen processing at a temperature of -20°C. A 10 K-type file was used for the initial collection of the samples followed by subsequent incubation of the files and paper pints in the incubation cabinet. Subsequent deoxyribonucleic acid (DNA) extraction from the samples was done following the procedure described by Siqueira et al. Paster et al's modification of the reverse-capture checkerboard assay was used in the present study. Semiquantitative data were used for overcoming the difficulties arising due to obtaining the counts of the polymerase chain reaction (PCR)-based analysis of specimens.Results: A positive result for the 16S ribosomal ribonucleic acid (rRNA) gene primer was observed only in two examined specimens of all the samples of the apical portion of the root canals in the present study. Negative result was shown by all the control group specimens, which were sterile samples. Presence of bacteria was confirmed by PCR in 38 out of 40 examined specimens. Amount of bacterial taxa, out of these 24 samples, ranged up to 6. Pseudoramibacter alactolyticus, Porphyromonas endodontalis, Dialister oral species, Bacteroidetes species, Streptococcus species, Olsenella uli, Synergistes species, Fusobacterium nucleatum, Parvimonas micra, Treponema denticola, and Filifactor alocis were the specific species detected. Bacteroidetes species was the only species that were detected at levels at or above 10 5 . Heavy bacterial infections were noticed in more than 45% of the cases at the periradicular part of the root canals. Conclusion:Microbial flora of the apical segment of the root with necroti...

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Selected Endodontic Pathogens in the Apical Third of Infected Root Canals: A Molecular Investigation

Cited by 65 publications

References 28 publications

Use of Multiple-Displacement Amplification and Checkerboard DNA-DNA Hybridization To Examine the Microbiota of Endodontic Infections

Use of Multiple-Displacement Amplification and Checkerboard DNA-DNA Hybridization To Examine the Microbiota of Endodontic Infections

Primer kok kanal enfeksiyonlarında Tannerella forsythia prevalansının 16S rRNA PCR ile incelenmesi

Evaluation of Bacteriological Profile in the Apical Root Segment of the Patients with Primary Apical Periodontitis

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