Multiplex Polymerase Chain Reaction Detection of Black-Pigmented Bacteria in Infections of Endodontic Origin

Seol, Jung-Hwan; Cho, Byung-Hoon; Chung, Chong-Pyoung; Bae, Kwang-Shik

doi:10.1016/j.joen.2005.10.020

Cited by 29 publications

(33 citation statements)

References 28 publications

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“…P. tannerae has been reported as an uncultivable organism (15), and its frequency of occurrence in endodontic infections was not appreciated until Xia et al (62) detected it in 60% of samples by use of PCR amplification. When multiplex PCR was used to detect BPB in endodontic samples, P. tannerae was found in only 5% of them, possibly due to limitations in the multiplex technique (38). Other BPB were also present in relatively high mean counts and propor-tions in amplified samples of the current study, including P. endodontalis, Prevotella loescheii, Prevotella nigrescens, Prevotella intermedia, P. gingivalis, and Prevotella melaninogenica.…”

Section: Discussionmentioning

confidence: 59%

“…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).…”

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confidence: 99%

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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: 59%

mentioning

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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“…1). Culture-independent molecular biology methods have allowed detection of this species in higher prevalence, ranging from 9%-72% (11)(12)(13)(14)(15)(16)(17)(18)(19)(20) (Fig. 1).…”

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confidence: 99%

Prevalence and Clonal Analysis of Porphyromonas gingivalis in Primary Endodontic Infections

Siqueira¹,

Rôças²,

Silva³

2008

Journal of Endodontics

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“…P. gingivalis and P. intermedia are black-pigmented gram-negative bacteria that colonize the subgingival crevice (18). P. gingivalis is consistently encountered in endodontic infections and is suspected to play a role in the etiology of acute abscesses (19). P. intermedia is among the most frequently detected species in primary caries infections (16).…”

Section: Discussionmentioning

confidence: 99%

Reactions of human dental pulp cells to capping agents in the presence or absence of bacterial exposure

Cai¹,

Zhang

Tribble

et al. 2017

J Oral Sci

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An ideal pulp-capping agent needs tohave good biocompatibility and promote reparative dentinogenesis. Although the effects of capping agents on healthy pulp are known, limited data regarding their effects on bacterial contaminated pulp are available. This study aimed to evaluate the reaction of contaminated pulps to various capping agents to assist clinicians in making informed decisions. Human dental pulp (HDP) cell cultures were developed from extracted human molars. The cells were exposed to a bacterial cocktail comprising Porphyromonas gingivalis, Prevotella intermedia, and Streptococcus gordonii before being cocultured with capping agents such as mineral trioxide aggregate (MTA) Portland cement (PC), and Dycal. HDP cell proliferation was assayed by MTS colorimetric cell proliferation assay, and its differentiation was evaluated by real-time PCR for detecting alkaline phosphatase, dentin sialophosphoprotein, and osteocalcin expressions. MTA and PC had no apparent effect, whereas Dycal inhibited HDP cell proliferation. PC stimulated HDP cell differentiation, particularly when they were exposed to bacteria. MTA and Dycal inhibited differentiation, regardless of bacterial infection. In conclusion, PC was the most favorable agent, followed by MTA, and Dycal was the least favorable agent for supporting the functions of bacterial compromised pulp cells.

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Multiplex Polymerase Chain Reaction Detection of Black-Pigmented Bacteria in Infections of Endodontic Origin

Cited by 29 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

Prevalence and Clonal Analysis of Porphyromonas gingivalis in Primary Endodontic Infections

Reactions of human dental pulp cells to capping agents in the presence or absence of bacterial exposure

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