Survival in transport media of <i>Actinobacillus actinomycetemcomitans, Porphymmonas gingivalis</i> and <i>Prevotella intermedia</i> in human subgingival samples

Steenbergen, T. J. M. van; Petit, Muriel; Tijhof, C. J.; Winkelhoff, A.J. van; Velden, U. van der; Graaff, J. de

doi:10.1111/j.1399-302x.1993.tb00613.x

Cited by 22 publications

(22 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…For this purpose, a P. gingivalis culture was serially diluted as described above. Paper points were incubated in the diluted culture until saturated and placed into reduced transport fluid (RTF)-containing vials (44). Four paper points per dilution were used.…”

Section: Bacteriamentioning

confidence: 99%

Highly Specific Protease-Based Approach for Detection of Porphyromonas gingivalis in Diagnosis of Periodontitis

et al. 2012

View full text Add to dashboard Cite

Porphyromonas gingivalis is associated with the development of periodontitis. Here we describe the development of a highly specific protease-based diagnostic method for the detection of P. gingivalis in gingival crevicular fluid. Screening of a proteolytic peptide substrate library, including fluorogenic dipeptides that contain D-amino acids, led to the discovery of five P. gingivalisspecific substrates. Due to the presence of lysine and arginine residues in these substrates, it was hypothesized that the cleavage was mediated by the gingipains, a group of P. gingivalis-specific proteases. This hypothesis was confirmed by the observation that P. gingivalis gingipain knockout strains demonstrated clearly impaired substrate cleavage efficacy. Further, proteolytic activity on the substrates was increased by the addition of the gingipain stimulators dithiothreitol and L-cysteine and decreased by the inhibitors leupeptin and N-ethylmaleimide. Screening of saliva and gingival crevicular fluid of periodontitis patients and healthy controls showed the potential of the substrates to diagnose the presence of P. gingivalis proteases. By using paper points, a sensitivity of approximately 10 5 CFU/ml was achieved. P. gingivalis-reactive substrates fully composed of L-amino acids and Bz-L-Arg-NHPhNO 2 showed a relatively low specificity (44 to 85%). However, the five P. gingivalis-specific substrates that each contained a single D-amino acid showed high specificity (96 to 100%). This observation underlines the importance of the presence of D-amino acids in substrates used for the detection of bacterial proteases. We envisage that these substrates may improve the specificity of the current enzyme-based diagnosis of periodontitis associated with P. gingivalis.

show abstract

Section: Bacteriamentioning

confidence: 99%

Highly Specific Protease-Based Approach for Detection of Porphyromonas gingivalis in Diagnosis of Periodontitis

et al. 2012

View full text Add to dashboard Cite

show abstract

“…VMGA III transport medium has been evaluated (3,8,46) specifically for anaerobic organisms associated with periodontal disease. VMGA III is a gel-based transport medium, and it has been argued that this results in maintenance of a low redox potential, thereby contributing to the enhanced survival of obligate anaerobes.…”

Section: Resultsmentioning

confidence: 99%

“…Canine periodontal pockets were sampled by veterinarians in France, Germany, Italy, Poland, Sweden, and the United Kingdom in the period of 2004 to 2006. Each veterinary practice was provided with sterile endodontic paper points for collection of periodontal pocket samples and airtight (screw-cap) polypropylene microcentrifuge tubes containing approximately 1.5 ml of sterile anaerobic VMGA III medium (46). The tubes were filled in an anaerobic workstation, and minimal headspace was left at the top of each tube prior to dispatch.…”

Section: Methodsmentioning

confidence: 99%

Activity of Pradofloxacin against Porphyromonas and Prevotella spp. Implicated in Periodontal Disease in Dogs: Susceptibility Test Data from a European Multicenter Study

Stephan

Greife

Pridmore

et al. 2008

Antimicrob Agents Chemother

View full text Add to dashboard Cite

Collaborating veterinarians from five European countries collected subgingival bacterial samples from dogs exhibiting clinical periodontal disease. Sterile endodontic paper points were used for collection of the samples, which were transported to a central laboratory for susceptibility testing. Anaerobic bacteria were isolated and Porphyromonas and Prevotella isolates identified to the species level; susceptibility to pradofloxacin and metronidazole was determined using the CLSI agar dilution methodology. A total of 630 isolates, 310 of Porphyromonas spp. and 320 of Prevotella spp., were isolated. Pradofloxacin MIC data for all isolates were in the range of <0.016 to 1 g/ml, the overall MIC 50 was 0.062, and the overall MIC 90 was 0.25 g/ml. There were no differences in activity against Porphyromonas and Prevotella isolates or in the pradofloxacin susceptibility distributions from the different European countries. All isolates were within the wild-type distribution and were fully susceptible to pradofloxacin. Metronidazole was also highly active against these strains: 316 of 320 Prevotella strains (98.8%) and 309 of 310 Porphyromonas strains (99.7%) were susceptible (MICs of <8 g/ml). However, three Prevotella strains had intermediate metronidazole susceptibility (MICs of 16 g/ml), while one Prevotella and one Porphyromonas strain were metronidazole resistant (MICs of 128 and 256 g/ml, respectively). Pradofloxacin, a novel broad-spectrum fluoroquinolone, demonstrates a high degree of antianaerobic activity against strains isolated from clinical cases of periodontal disease and shows activity against metronidazole-resistant isolates. The broad-spectrum activity of pradofloxacin makes it a suitable candidate for the treatment of periodontal disease in dogs.

show abstract

“…The QC specimen (approximately 10 10 cfu/mL) was prepared in thioglycolate broth of which 100 µl was transferred into anaerobically prepared 2 ml VMGAIII transport medium vials (4) and sent to participating laboratories by a courier. The specimen was accompanied with the following information: ‘The specimen is from a 7 mm periodontal pocket, upper 1 st molar in a 56-year old male patient with type II diabetes and is allergic to penicillin’.…”

Section: Methodsmentioning

confidence: 99%

Quality control for diagnostic oral microbiology laboratories in European countries

Rautemaa‐Richardson

Reijden

Dahlén

et al. 2011

Journal of Oral Microbiology

View full text Add to dashboard Cite

Participation in diagnostic microbiology internal and external quality control (QC) processes is good laboratory practice and an essential component of a quality management system. However, no QC scheme for diagnostic oral microbiology existed until 2009 when the Clinical Oral Microbiology (COMB) Network was created. At the European Oral Microbiology Workshop in 2008, 12 laboratories processing clinical oral microbiological samples were identified. All these were recruited to participate into the study and six laboratories from six European countries completed both the online survey and the first QC round. Three additional laboratories participated in the second round. Based on the survey, European oral microbiology laboratories process a significant (mean per laboratory 4,135) number of diagnostic samples from the oral cavity annually. A majority of the laboratories did not participate in any internal or external QC programme and nearly half of the laboratories did not have standard operating procedures for the tests they performed. In both QC rounds, there was a large variation in the results, interpretation and reporting of antibiotic susceptibility testing among the laboratories. In conclusion, the results of this study demonstrate the need for harmonisation of laboratory processing methods and interpretation of results for oral microbiology specimens. The QC rounds highlighted the value of external QC in evaluating the efficacy and safety of processes, materials and methods used in the laboratory. The use of standardised methods is also a prerequisite for multi-centre epidemiological studies that can provide important information on emerging microbes and trends in anti-microbial susceptibility for empirical prescribing in oro-facial infections.

show abstract

Survival in transport media of Actinobacillus actinomycetemcomitans, Porphymmonas gingivalis and Prevotella intermedia in human subgingival samples

Cited by 22 publications

References 15 publications

Highly Specific Protease-Based Approach for Detection of Porphyromonas gingivalis in Diagnosis of Periodontitis

Highly Specific Protease-Based Approach for Detection of Porphyromonas gingivalis in Diagnosis of Periodontitis

Activity of Pradofloxacin against Porphyromonas and Prevotella spp. Implicated in Periodontal Disease in Dogs: Susceptibility Test Data from a European Multicenter Study

Quality control for diagnostic oral microbiology laboratories in European countries

Contact Info

Product

Resources

About