Rapid presumptive identification of black-pigmented gram-negative anaerobic bacteria by using 4-methylumbelliferone derivatives

Moncla, Bernard J.; Braham, Pamela H.; Rabe, Lorna K.; Hillier, Sharon L.

doi:10.1128/jcm.29.9.1955-1958.1991

Cited by 55 publications

(19 citation statements)

References 20 publications

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“…MLEE provides a definitive distinction between P. intermedia and P. nigrescens [1] but MDH and GDH enzymes from the vaginal isolates did not have migration rates which corresponded to either species. The reactivity of the three strains with the mAbs used here also indicates that they are not P. intermedia or P. nigrescens, and together these results strengthen the proposal made by Moncla et al [16] that they represent a previously undescribed species which is nonetheless biochemically similar to P. intermedia and P. nigrescens. Further studies of these and other organisms identified as P. intermedia, and isolated from non-oral sites, are in progress.…”

Section: Discussionsupporting

confidence: 88%

“…The vaginal isolates used in this study were shown previously to be atypical in that they failed to react with RNA probes prepared against nucleic acid of strains of genotypes I or II of P. interrnedia (i.e.P. intermedia and P. nigrescens), and they were a-fucosidase negative [16]. MLEE provides a definitive distinction between P. intermedia and P. nigrescens [1] but MDH and GDH enzymes from the vaginal isolates did not have migration rates which corresponded to either species.…”

Section: Discussionmentioning

confidence: 95%

“…Nine isolates (prefix MH) were provided by M. Haapasaalo (Department of Cariology, University of Helsinki, Finland) and were from endodontic infections and periodontal pockets. HST 1156-3, HST 2160-1 and HST 2166-2 were provided by P. Braham (Research Centre in Oral Biology, University of Washington, USA); these were isolated from vaginal sites and were identified biochemically and by gas liquid chromatography as P. intermedia, but they were a-fucosidase negative and did not hybridize with nucleic acid probes raised against P. intermedia genotypes I and II [16].…”

Section: Bacterial Strainsmentioning

confidence: 99%

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Species-specificity of monoclonal antibodies recognising Prevotella intermedia and Prevotella nigrescens

Devine¹

1994

FEMS Microbiology Letters

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Prevotella intermedia and Prevotella nigrescens are not easily distinguished, making it difficult to assess their roles in disease. This study examined the specificity of three monoclonal antibodies (mAbs) for these species. Differentiation between P. intermedia (13 isolates) and P. nigrescens (24 isolates) was by the electrophoretic mobility of their malate and glutamate dehydrogenase enzymes or by DNA homology grouping. All P. intermedia reacted strongly with mAb 40BI3.2.2 whereas P. nigrescens strains did not. Monoclonal antibodies 37BI6.1 and 39BI1.1.2 recognised all strains of both species but most P. nigrescens reacted weakly with mAb 39BI1.1.2. Monoclonal antibody 40BI3.2.2 therefore recognises an antigen specific for P. intermedia but not P. nigrescens and provides an easy and reliable means of distinguishing between these species. Three vaginal isolates identified biochemically as P. intermedia had enzymes with mobilities corresponding to neither P. intermedia nor P. nigrescens. These isolates were not recognised by mAbs 39BI1.1.2 or 40BI3.2.2 and may represent an undescribed taxon within this group of organisms.

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Section: Discussionsupporting

confidence: 88%

Section: Discussionmentioning

confidence: 95%

Section: Bacterial Strainsmentioning

confidence: 99%

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Species-specificity of monoclonal antibodies recognising Prevotella intermedia and Prevotella nigrescens

Devine¹

1994

FEMS Microbiology Letters

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“…The chnical isolates were designated according to patient number, site number and isolate number as follows: P 4.3.5, P 6. All isolates were identified according to the methods of Moncla et al (12). Five commonly used laboratory isolates (W50, W83, ATCC 49417, Pg22 KNG.12, PgIET3.10) were also included in the study.…”

Section: Materials and Methods Bacterial Strains And Culture Conditionsmentioning

confidence: 99%

Natural variation within the principal arginine‐specific protease gene, prpR1, of Porphyromonas gingivalis

Allaker¹,

Aduse-Opoku²,

Batten³

et al. 1997

Oral Microbiology and Immunology

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RI, one of the major extracellular arginine-specific proteases of Porphyromonas gingivalis is a heterodimer composed of catalytic (alpha) and adhesin (beta) chains, encoded by the gene prpR1. The distribution of prpR1 and its variation within 43 isolates of P. gingivalis was determined. Chromosomal DNA was digested with Sma I and probed with a 32P-labeled DNA fragment from within the coding region for the alpha component of P. gingivalis W50. All isolates gave the expected 3.2 kb band, corresponding to the coding region for the alpha and beta components. The presence of a second locus (prR2) homologous to the alpha region of prpR1 was also detected. The 1.7-kb alpha coding region of prpR1 was amplified for subsequent restriction analysis. Following Taq I restriction all isolates gave identical patterns. With Rsa I, the majority of isolates (77%) could be placed into a single group. In conclusion, the prpR1 and prR2 loci are maintained in natural populations of P. gingivalis, and only minor polymorphism is detectable within the catalytic domain.

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“…All P. gingivalis strains were originally human isolates donated to the laboratory. Species identity was confirmed via reactivity with a range of fluorescent substrates for hydrolytic enzymes basically as described by Moncla et al [20] except a 96-well microtitire plate was used for assays. Bacteria were grown in brain-heart infusion broth (Oxoid, Basingstoke, UK) supplemented with 5 mg/1 hemin.…”

Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

Production and characterisation of monoclonal antibodies to the principle sonicate antigens of Porphyromonas gingivalis w50

Millar

1993

FEMS Immunology and Medical Microbiology

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Protein antigens from whole cell sonicates of Porphyromonas gingivalis W50, previously shown to be discriminatory antigens for patients with adult periodontitis, were purified using SDS-PAGE. Electroeluted proteins were used to immunize mice for the production of monoclonal antibodies (mAbs). A combination of enzyme-linked immunosorbent assay (ELISA) and Western blotting were used to screen hybridoma supernatants for mAbs. MAbs were successfully raised against M(r) 115,000, M(r) 55,000 and M(r) 47,000 antigens together with a second M(r) 55,000 polypeptide which was a contaminant of the M(r) 55,000 antigen. No immunological cross-reactivity was found between these four proteins. The mAbs were used to examine the distribution of these antigens among fifteen P. gingivalis strains together with related oral bacteria using immunostaining of dot blots and Western blots. The antigens were confined to P. gingivalis with the M(r) 115,000 and M(r) 47,000 antigens being present in all strains tested. The distribution of the M(r) 55,000 antigens were slightly more restricted: one M(r) 55,000 (outer membrane location) was present in nine of the fifteen P. gingivalis strains tested, while the other M(r) 55,000 (location unknown) was only absent from one strain. Whole cell ELISA demonstrated that the M(r) 115,000 and the outer membrane M(r) 55,000 antigen possess epitopes which are located on the surface of the bacterium.

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Rapid presumptive identification of black-pigmented gram-negative anaerobic bacteria by using 4-methylumbelliferone derivatives

Cited by 55 publications

References 20 publications

Species-specificity of monoclonal antibodies recognising Prevotella intermedia and Prevotella nigrescens

Species-specificity of monoclonal antibodies recognising Prevotella intermedia and Prevotella nigrescens

Natural variation within the principal arginine‐specific protease gene, prpR1, of Porphyromonas gingivalis

Production and characterisation of monoclonal antibodies to the principle sonicate antigens of Porphyromonas gingivalis w50

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