Open Access, Peer-reviewed
eISSN 2234-7666
    J Korean Acad Conserv Dent. 2002 Jan;27(1):54-65. Korean.
    Published online Jan 31, 2002.
    https://doi.org/10.5395/JKACD.2002.27.1.054
    Copyright © 2002 Korean Academy of Conservative Dentistry
    Original Article

    Detection of black-pigmented bacteria in infected root canals

    Eun-Kyoung Kwon, Eun-Sook Kim, Ju-Seog Kwak, Hwang Lee, Su-Jong Lee and Mi-Kyung Im
      • Department of Conservative Dentistry, College of Dentistry, Wonkwang University, Korea.

    This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

    Abstract

    Black-pigmented bacteria have been implicated in the endodontic infections. This group of microorganisms includes Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens. The organisms display a wide variety of virulence factors that may be pertinent to acute endodontic infections.

    The aim of this study was to identify P. endodontalis, P. gingivalis, P. intermedia, and P. nigrescens by using special potency disk test, filter paper spot test, 16S rRNA gene-directed PCR, and API 32A.

    Microbial samples were collected from root canals of 33 intact teeth with necrotic pulp and/or apical periodontitis. Conventional laboratory methods were used for identification of the strains of black pigmented bacteria. Eighteen of 33 samples were positive for the growth of black-pigmented bacrteria. Five colonies were cultured from each pure cultured colonies from Brucella agar plate. Seventy seven colonies were positive for the growth of black-pigmented bacteria.

    Thirty three of 77(42.6%) were identifed as P. nigrescens, 10 of 77(12.9%) were P. gingivalis, 6 of 77(7.8%) were P. endodontalis, 10 of 77(12.9%) were P. intermedia. On the contrary the reference strains of P. nigrescens, experimental strains of P. nigrescens was sensitive to kanamycin in special potency disk test.

    16S rRNA gene PCR and API test after rapid presumptative identification methods, such as special potency disk test and filter paper spot test, would be accurate detection methods for black-pigemented bacteria.

    Figures

    Fig. 1
    Primary culture sample

    Fig. 2
    Specific potency disk test of P. gingivalis(ATCC 33277)

    P. gingivalis was resistant to kanamycin and colistin, and sensitive to vancomycin.

    Fig. 3
    Specific disk potency test of P. intermedia(ATCC 25611)

    P. intermedia was resistant to kanamycin and vancomycin, and sensitive to colistin.

    Fig. 4
    Specific potency disk test of P. nigrescens(ATCC 33563)

    P. nigrescens was resistant to kanamycin and vancomycin, and sensitive to colistin.

    Fig. 5
    Special potency disk test of patient sample was identified P. nigrescens.

    P. nigrescens was resistant to vancomycin, and sensitive to kanamycin and colistin.

    Fig. 6
    Filter paper spot test result of patient sample was identified P. nigrescens.

    from left to right: trypsin like enzyme, α-glucosidase, β-glutosidase, α-fucosidase P. nigrescens showed positive response to α-glucosidase and α-fucosidase.

    Fig. 7
    PCR profiles of P. endodontalis, P. gingivalis, P. nigrescens, P. intermedia amplified with species specific primers for the 16S rRNA

    SM: size marker(1kb DNA ladder)

    lane 1: P. endodontalis(ATCC 35406)

    lane 2: P. gingivalis(ATCC 33277)

    lane 3: P. nigrescens(ATCC 33563)

    lane 4: P. intermedia(ATCC 25611)

    Fig. 8
    Agarose gel electrophoresis patterns of PCR products of samples

    SM: size marker(1kb DNA ladder)

    lane 1: sample No. 201(P. nigrescens)

    lane 2: sample No. 2803(P. intermedia)

    lane 3: sample No. 2806(P. intermedia)

    lane 4: sample No. 2401(P. gingivalis)

    lane 5: sample No. 2403(P. gingivalis)

    lane 6: sample No. 3201(P. nigrescens)

    lane 7: sample No. 3205(P. nigrescens)

    Fig. 9
    RapidID 32A test result of clinical isolates

    Tables

    Table 1
    Special potency disk test on ATCC type strains

    Table 2
    Special potency disk test on clinical isolates

    Table 3
    Filter paper spot test on ATCC type strains

    Table 4
    Filter paper spot test on clinical isolates

    Table 5
    Number of strains clinical isolates identified using PCR

    Table 6
    Number of strains clinical isolates identified using RapidID 32A

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    MeSH Terms
    Agar
    Bacteria
    Brucella
    Dental Pulp Cavity
    Genes, rRNA
    Kanamycin
    Periapical Periodontitis
    Polymerase Chain Reaction
    Porphyromonas endodontalis
    Porphyromonas gingivalis
    Prevotella intermedia
    Prevotella nigrescens
    Tooth
    Virulence Factors
    Metrics
    Share
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    Tables
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