Logo of jcmPermissionsJournals.ASM.orgJournalJCM ArticleJournal InfoAuthorsReviewers
J Clin Microbiol. Feb 1996; 34(2): 376–384.
PMCID: PMC228801

Rapid characterization of periodontal bacterial isolates by using fluorogenic substrate tests.

Abstract

Eighty-nine species of subgingival bacteria, represented by 121 reference strains and 892 patient isolates, including gram-negative, gram-positive, aerobic, facultatively anaerobic, microaerophilic, and anaerobic species, were characterized with a panel of fluorogenic, 4-methylumbelliferyl-linked substrate tests. Identifications of all patient isolates were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of whole-cell proteins relative to reference strains. Characteristic profiles of positive fluorogenic reactions differentiated most of the species, including five Porphyromonas species, six pigmenting and five nonpigmenting Prevotella species, Bacteroides forsythus, three Capnocytophaga species, six Actinomyces species, four Propionibacterium species, and eight Streptococcus species. Two mannoside isomers differentiated Actinomyces israelii and Actinomyces gerencseriae. In addition to Porphyromonas gingivalis, B. forsythus, and Capnocytophaga species, Fusobacterium alocis, Actinomyces odontolyticus, Actinomyces meyeri, and Bifidobacterium dentium were all positive for so-called trypsin-like activity. Fusobacterium nucleatum, Eikenella corrodens, Actinobacillus actinomycetemcomitans, and Campylobacter species were nonreactive with the carbohydrate-based substrates tested. Fluorogenic substrate tests provided a sensitive and simple method for biochemical characterization that could presumptively identify to species level most subgingival isolates within 4 h. The method was ideal for rapidly obtaining presumptive identifications of isolates prior to confirming identifications by definitive methods, such as SDS-PAGE.

Full Text

The Full Text of this article is available as a PDF (245K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aicoforado GA, McKay TL, Slots J. Rapid method for detection of lactose fermenting oral microorganisms. Oral Microbiol Immunol. 1987 Mar;2(1):35–38. [PubMed]
  • Bedi GS, Williams T. Purification and characterization of a collagen-degrading protease from Porphyromonas gingivalis. J Biol Chem. 1994 Jan 7;269(1):599–606. [PubMed]
  • Beighton D, Hardie JM, Whiley RA. A scheme for the identification of viridans streptococci. J Med Microbiol. 1991 Dec;35(6):367–372. [PubMed]
  • Bruun B, Ying Y, Kirkegaard E, Frederiksen W. Phenotypic differentiation of Cardiobacterium hominis, Kingella indologenes and CDC group EF-4. Eur J Clin Microbiol. 1984 Jun;3(3):230–235. [PubMed]
  • Collins MD, Love DN, Karjalainen J, Kanervo A, Forsblom B, Willems A, Stubbs S, Sarkiala E, Bailey GD, Wigney DI, et al. Phylogenetic analysis of members of the genus Porphyromonas and description of Porphyromonas cangingivalis sp. nov. and Porphyromonas cansulci sp. nov. Int J Syst Bacteriol. 1994 Oct;44(4):674–679. [PubMed]
  • Dellinger CA, Moore LV. Use of the RapID-ANA System to screen for enzyme activities that differ among species of bile-inhibited Bacteroides. J Clin Microbiol. 1986 Feb;23(2):289–293. [PMC free article] [PubMed]
  • Elliott JA, Facklam RR. Identification of Leuconostoc spp. by analysis of soluble whole-cell protein patterns. J Clin Microbiol. 1993 May;31(5):1030–1033. [PMC free article] [PubMed]
  • Heukeshoven J, Dernick R. Increased sensitivity for Coomassie staining of sodium dodecyl sulfate-polyacrylamide gels using PhastSystem Development Unit. Electrophoresis. 1988 Jan;9(1):60–61. [PubMed]
  • Hinnebusch CJ, Nikolai DM, Bruckner DA. Comparison of API Rapid Strep, Baxter MicroScan Rapid Pos ID Panel, BBL Minitek Differential Identification System, IDS RapID STR System, and Vitek GPI to conventional biochemical tests for identification of viridans streptococci. Am J Clin Pathol. 1991 Oct;96(4):459–463. [PubMed]
  • Johnson JL, Moore LV, Kaneko B, Moore WE. Actinomyces georgiae sp. nov., Actinomyces gerencseriae sp. nov., designation of two genospecies of Actinomyces naeslundii, and inclusion of A. naeslundii serotypes II and III and Actinomyces viscosus serotype II in A. naeslundii genospecies 2. Int J Syst Bacteriol. 1990 Jul;40(3):273–286. [PubMed]
  • Khwaja KJ, Parish P, Aldred MJ, Wade WG. Protein profiles of Capnocytophaga species. J Appl Bacteriol. 1990 Apr;68(4):385–390. [PubMed]
  • Kilian M. Rapid identification of Actinomycetaceae and related bacteria. J Clin Microbiol. 1978 Aug;8(2):127–133. [PMC free article] [PubMed]
  • Kinder SA, Kornman KS, Holt SC. Characterization of selected gram-negative oral microorganisms by SDS-PAGE. Oral Microbiol Immunol. 1989 Mar;4(1):52–56. [PubMed]
  • Lala A, Amano A, Sojar HT, Radel SJ, De Nardin E. Porphyromonas gingivalis trypsin-like protease: a possible natural ligand for the neutrophil formyl peptide receptor. Biochem Biophys Res Commun. 1994 Mar 30;199(3):1489–1496. [PubMed]
  • Laughon BE, Syed SA, Loesche WJ. API ZYM system for identification of Bacteroides spp., Capnocytophaga spp., and spirochetes of oral origin. J Clin Microbiol. 1982 Jan;15(1):97–102. [PMC free article] [PubMed]
  • Love DN, Bailey GD, Collings S, Briscoe DA. Description of Porphyromonas circumdentaria sp. nov. and reassignment of Bacteroides salivosus (Love, Johnson, Jones, and Calverley 1987) as Porphyromonas (Shah and Collins 1988) salivosa comb. nov. Int J Syst Bacteriol. 1992 Jul;42(3):434–438. [PubMed]
  • Maiden MF, Tanner A. Identification of oral yeasts by polyacrylamide gel electrophoresis. Oral Microbiol Immunol. 1991 Jun;6(3):187–190. [PubMed]
  • Maiden MF, Tanner A, Moore WE. Identification of Selenomonas species by whole-genomic DNA probes, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, biochemical tests and cellular fatty acid analysis. Oral Microbiol Immunol. 1992 Feb;7(1):7–13. [PubMed]
  • Manafi M, Kneifel W. Rapid methods for differentiating gram-positive from gram-negative aerobic and facultative anaerobic bacteria. J Appl Bacteriol. 1990 Dec;69(6):822–827. [PubMed]
  • Manafi M, Kneifel W, Bascomb S. Fluorogenic and chromogenic substrates used in bacterial diagnostics. Microbiol Rev. 1991 Sep;55(3):335–348. [PMC free article] [PubMed]
  • Manafi M, Rotter ML. A new plate medium for rapid presumptive identification and differentiation of Enterobacteriaceae. Int J Food Microbiol. 1991 Nov;14(2):127–134. [PubMed]
  • Millership SE, Want SV. Whole-cell protein electrophoresis for typing Mycobacterium tuberculosis. J Clin Microbiol. 1992 Nov;30(11):2784–2787. [PMC free article] [PubMed]
  • Moncla BJ, Braham P. Detection of sialidase (neuraminidase) activity in Actinomyces species by using 2'-(4-methylumbelliferyl)alpha-D-N-acetylneuraminic acid in a filter paper spot test. J Clin Microbiol. 1989 Jan;27(1):182–184. [PMC free article] [PubMed]
  • Moncla BJ, Braham P, Hillier SL. Sialidase (neuraminidase) activity among gram-negative anaerobic and capnophilic bacteria. J Clin Microbiol. 1990 Mar;28(3):422–425. [PMC free article] [PubMed]
  • Moncla BJ, Braham P, Rabe LK, Hillier SL. Rapid presumptive identification of black-pigmented gram-negative anaerobic bacteria by using 4-methylumbelliferone derivatives. J Clin Microbiol. 1991 Sep;29(9):1955–1958. [PMC free article] [PubMed]
  • Moore LV, Moore WE. Oribaculum catoniae gen. nov., sp. nov.; Catonella morbi gen. nov., sp. nov.; Hallella seregens gen. nov., sp. nov.; Johnsonella ignava gen. nov., sp. nov.; and Dialister pneumosintes gen. nov., comb. nov., nom. rev., Anaerobic gram-negative bacilli from the human gingival crevice. Int J Syst Bacteriol. 1994 Apr;44(2):187–192. [PubMed]
  • Moore LV, Johnson JL, Moore WE. Descriptions of Prevotella tannerae sp. nov. and Prevotella enoeca sp. nov. from the human gingival crevice and emendation of the description of Prevotella zoogleoformans. Int J Syst Bacteriol. 1994 Oct;44(4):599–602. [PubMed]
  • Moore WE, Hash DE, Holdeman LV, Cato EP. Polyacrylamide slab gel electrophoresis of soluble proteins for studies of bacterial floras. Appl Environ Microbiol. 1980 Apr;39(4):900–907. [PMC free article] [PubMed]
  • Paster BJ, Dewhirst FE, Olsen I, Fraser GJ. Phylogeny of Bacteroides, Prevotella, and Porphyromonas spp. and related bacteria. J Bacteriol. 1994 Feb;176(3):725–732. [PMC free article] [PubMed]
  • Pederson ED, Miller JW, Matheson S, Simonson LG, Chadwick DE, Covill PJ, Turner DW, Lamberts BL, Morton HE. Trypsin-like activity levels of Treponema denticola and Porphyromonas gingivalis in adults with periodontitis. J Clin Periodontol. 1994 Sep;21(8):519–525. [PubMed]
  • Pot B, Hertel C, Ludwig W, Descheemaeker P, Kersters K, Schleifer KH. Identification and classification of Lactobacillus acidophilus, L. gasseri and L. johnsonii strains by SDS-PAGE and rRNA-targeted oligonucleotide probe hybridization. J Gen Microbiol. 1993 Mar;139(3):513–517. [PubMed]
  • Slayne MA, Aldred MJ, Wade WG. A rapid, semi-automated SDS-PAGE identification system for oral anaerobic bacteria. J Appl Bacteriol. 1990 Apr;68(4):391–395. [PubMed]
  • Slots J. Enzymatic characterization of some oral and nonoral gram-negative bacteria with the API ZYM system. J Clin Microbiol. 1981 Sep;14(3):288–294. [PMC free article] [PubMed]
  • Slots J. Detection of colonies of Bacteroides gingivalis by a rapid fluorescence assay for trypsin-like activity. Oral Microbiol Immunol. 1987 Sep;2(3):139–141. [PubMed]
  • Tanner A, Maiden MF, Paster BJ, Dewhirst FE. The impact of 16S ribosomal RNA-based phylogeny on the taxonomy of oral bacteria. Periodontol 2000. 1994 Jun;5:26–51. [PubMed]
  • Tanner AC. Characterization of Wolinella spp., Campylobacter concisus, Bacteroides gracilis, and Eikenella corrodens by polyacrylamide gel electrophoresis. J Clin Microbiol. 1986 Oct;24(4):562–565. [PMC free article] [PubMed]
  • Tanner AC, Strzempko MN, Belsky CA, McKinley GA. API ZYM and API An-Ident reactions of fastidious oral gram-negative species. J Clin Microbiol. 1985 Sep;22(3):333–335. [PMC free article] [PubMed]
  • Vandamme P, Falsen E, Pot B, Hoste B, Kersters K, De Ley J. Identification of EF group 22 campylobacters from gastroenteritis cases as Campylobacter concisus. J Clin Microbiol. 1989 Aug;27(8):1775–1781. [PMC free article] [PubMed]
  • van Winkelhoff AJ, Clement M, de Graaff J. Rapid characterization of oral and nonoral pigmented Bacteroides species with the ATB Anaerobes ID system. J Clin Microbiol. 1988 May;26(5):1063–1065. [PMC free article] [PubMed]
  • van Winkelhoff AJ, van Steenbergen TJ, Kippuw N, de Graaff J. Enzymatic characterization of oral and non-oral black-pigmented Bacteroides species. Antonie Van Leeuwenhoek. 1986;52(2):163–171. [PubMed]
  • Whiley RA, Fraser H, Hardie JM, Beighton D. Phenotypic differentiation of Streptococcus intermedius, Streptococcus constellatus, and Streptococcus anginosus strains within the "Streptococcus milleri group". J Clin Microbiol. 1990 Jul;28(7):1497–1501. [PMC free article] [PubMed]
  • Whiley RA, Fraser HY, Douglas CW, Hardie JM, Williams AM, Collins MD. Streptococcus parasanguis sp. nov., an atypical viridans Streptococcus from human clinical specimens. FEMS Microbiol Lett. 1990 Mar 1;56(1-2):115–121. [PubMed]
  • Whiley RA, Hardie JM. DNA-DNA hybridization studies and phenotypic characteristics of strains within the 'Streptococcus milleri group'. J Gen Microbiol. 1989 Oct;135(10):2623–2633. [PubMed]
  • Willems A, Collins MD. Reclassification of Oribaculum catoniae (Moore and Moore 1994) as Porphyromonas catoniae comb. nov. and emendation of the genus Porphyromonas. Int J Syst Bacteriol. 1995 Jul;45(3):578–581. [PubMed]
  • Willems A, Collins MD. Phylogenetic placement of Dialister pneumosintes (formerly Bacteroides pneumosintes) within the Sporomusa subbranch of the Clostridium subphylum of the gram-positive bacteria. Int J Syst Bacteriol. 1995 Apr;45(2):403–405. [PubMed]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...