Biomedical subjects

T F Flemmig

Publications and source records attributed to T F Flemmig.

At least 37 records · Page 2Linked to original sources

PubMed1999

Simultaneous detection of Bacteroides forsythus and Prevotella intermedia by 16S rRNA gene-directed multiplex PCR.

In a 16S rRNA gene-directed multiplex PCR, Prevotella intermedia- and Bacteroides forsythus-specific reverse primers were combined with a single conserved forward primer. A 660-bp fragment and an 840-bp fragment that were specific for both species could be amplified simultaneously. A total of 152 clinical samples, subgingival plaque and swabs of three different oral mucosae, from 38 periodontitis patients were used for the evaluation.

Adult↗

PubMed1999

Clonal infection with Actinobacillus actinomycetemcomitans following periodontal therapy.

Mechanical debridement results in a shift of the bacterial composition in the periodontal pocket on the species level. It is unknown, however, whether a clonal change within a species could lead to the emergence of strains with different levels of virulence. Therefore, in the present study, the genetic variability of Actinobacillus actinomycetemcomitans was assessed and strains identified which were associated with periodontal disease progression following periodontal therapy, i.e., refractory periodontitis. Twenty adult patients with untreated periodontitis and subgingival colonization of A. actinomycetemcomitans were randomly assigned to receive full-mouth scaling alone or scaling with an adjunctive antimicrobial therapy. Both groups received supportive periodontal therapy at 3, 6, 9, 12, 18, and 24 months. Subgingival plaque samples were taken at every visit; venous blood was obtained at 24 months only. A. actinomycetemcomitans isolates were typed by the RAPD method, and antibody reactivity against outer membrane proteins was assessed by immunoblot analysis. Eleven distinct RAPD patterns were found in 18 patients completing the study. All patients harbored only one A. actinomycetemcomitans genotype, and within each patient this genotype persisted throughout the 24-month observation period. No differences in the expression of antibody reactivity against outer membrane proteins were found between strains isolated at baseline and at 24 months. Three genotypes were associated with reduced survival rates of teeth without probing attachment loss of 2 mm or more. The results indicated that (i) most patients harbored only one A. actinomycetemcomitans genotype; (ii) the genotype persisted following therapy; and (iii) only some genotypes were associated with refractory periodontitis.

Adult↗

PubMed1999

Periodontitis.

The purpose of this review was to assess the scientific and clinical bases for the proposed classification of periodontitis. The clinical and histopathological signs and the etiology of periodontitis were described. Cross-sectional studies were analyzed to determine when onset of periodontitis most frequently occurs in adults. In addition, the progression rates of periodontitis have been assessed from longitudinal studies. No clinical, histopathological, or microbiological features could be identified that would characterize different disease entities of chronic periodontitis. The prevalence, extent, and severity of periodontitis were found to increase continually with higher age and there was no age when onset of disease would most likely occur. The rate of periodontitis progression varies largely between patients and there is no natural threshold for distinguishing various rates of disease progression. The incidence of periodontitis unresponsive to treatment depends on pretreatment progression rate, extent and severity of disease, tooth type, smoking, high levels of putative periodontal pathogens, a deficient immune response, and the type of therapy provided. There is no scientific basis for the classification "adult periodontitis" and "refractory adult periodontitis." Extensive clinical examinations are required for the diagnosis of "rapidly progressive adult periodontitis." It appears unrealistic that these examinations can be performed routinely in clinical practice. Therefore, the classification proposed by the Organizing Committee to define adult, rapidly progressive, and refractory periodontitis as specific disease entities was replaced with a simplified classification of periodontitis based on the scientific data available.

Acute Disease↗

PubMed1999

Combined systemic and local antimicrobial therapy of periodontal disease in Papillon-Lefèvre syndrome. A report of 4 cases.

4 patients, 2 pairs of siblings, suffering from Papillon-Lefèvre syndrome were treated for periodontal disease. Following extraction of hopeless teeth, the children received scaling and adjunctive systemic antibiotics (metronidazole and amoxicillin for 7 to 10 days). In addition, they performed supragingival pulsated jet irrigation with 0.06% chlorhexidine digluconate 1 x daily. In 2 siblings, A. actinomycetemcomitans was suppressed subgingivally below detectable levels, pocket probing depths were reduced to 4 mm or less, and plaque and bleeding indices were low. No further disease progression was seen over a 3-year-period. Another female patient also showed clinical improvement and suppression of subgingival A. actinomycetemcomitans and B. forsythus up to the 9-month-follow-up, while her sister showed further attachment loss over the course of 4 years. The present case reports indicated that in some patients suffering from Papillon-Lefèvre syndrome periodontal disease may be arrested by means of (i) oral hygiene instruction, (ii) extraction of severely diseased teeth, (iii) scaling, (iv) systemic antibiotics and (v) long-term antimicrobial irrigation.

Aggregatibacter actinomycetemcomitans↗

PubMed1998

Differential effects of systemic metronidazole and amoxicillin on Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in intraoral habitats.

48 adult patients with untreated periodontitis harboring subgingival Actinobacillus actinomycetemcomitans and/or Porphyromonas gingivalis were randomly assigned to receive full mouth scaling alone (control) or scaling with systemic metronidazole plus amoxicillin and supragingivai irrigation with chlorhexidine digluconate (test). Subgingival plaque and swab samples from tongue, tonsils, and buccal mucosa were taken at baseline, 10 days and 3, 6, 9, and 12 months. A. actinomycetemcomitans was detected in the oral cavity, i.e., subgingival plaque and/or mucous membranes, less frequently in test patients compared to controls at 9 and 12 months (p<0.01), whereas, the intraoral detection frequency of P. gingivalis was significantly reduced only 10 days following therapy (p<0.001). At any time after therapy, A. actinomycetemcomitans was not detected intraorally in 5 of 10 (50%) test and 1 of 13 (8%) control patients harboring this pathogen at baseline; P. gingivalis was not detected in only 1 of 18 (6%) test and none of the 17 control patients harboring this pathogen at baseline. Although the data indicated that the assessed antimicrobial therapy may suppress A. actinomycetemcomitans from the entire oral cavity below detectable levels over a minimum of 12 months, P. gingivalis persisted or reoccurred.

Administration, Oral↗

PubMed1998

The effect of working parameters on root substance removal using a piezoelectric ultrasonic scaler in vitro.

This study assessed defect depth and volume resulting from root instrumentation using a piezoelectric ultrasonic scaler with a slim scaling tip in vitro. Combinations of the following working parameters were analyzed: lateral forces of 0.5 N, 1 N, and 2 N; tip angulations of 0 degrees, 45 degrees, and 90 degrees; power settings of low, medium and high; and instrumentation time of 10 s, 20 s, 40 s, and 80 s. Defects were quantified using a 3D optical laser scanner. Overall, lateral force had the greatest influence on defect volume compared to instrument power setting and tip angulation (beta-weights 0.49 +/- 0.04, 0.25 +/- 0.04, and 0.14 +/- 0.04, respectively). The effects on defect depth were highest for tip angulation followed by lateral force and instrument power setting (beta-weights 0.48 +/- 0.04, 0.34 +/- 0.04, and 0.25 +/- 0.04, respectively). Interestingly, at all power settings, the highest defect volume and depth by far were found after combining 45 degrees tip angulation with 2 N of lateral force. The efficacy of the assessed piezoelectric ultrasonic scaler may be adapted to the various clinical needs by adjusting the lateral force, tip angulation, and power setting. To prevent severe root damage it is crucial to use the assessed scaler at a tip angulation of close to 0 degrees.

Dental Instruments↗

PubMed1998

Differential clinical treatment outcome after systemic metronidazole and amoxicillin in patients harboring Actinobacillus actinomycetemcomitans and/or Porphyromonas gingivalis.

48 adult patients with untreated periodontitis harboring subgingival Actinobacillus actinomycetemcomitans and/or Porphyromonas gingivalis as assessed by PCR were randomly assigned to receive full-mouth scaling alone (control) or scaling with systemic metronidazole plus amoxicillin and supragingival irrigation with chlorhexidine digluconate (test). In patients harboring A. actinomycetemcomitans intraorally at baseline, the adjunctive antimicrobial therapy resulted in a significantly higher incidence of probing attachment level (PAL) gain of 2 mm or more compared to scaling alone over 12 months (p<0.05). In addition, suppression of A. actinomycetemcomitans in subgingival plaque below detectable levels was associated with an increased incidence of PAL gain. In contrast, patients initially harboring P. gingivalis but not A. actinomycetemcomitans in the oral cavity showed a significantly higher incidence of PAL loss following adjunctive antimicrobial therapy compared to scaling alone (p<0.05). When the presence of pathogens at baseline was disregarded in the analysis, adjunctive antimicrobial therapy did not significantly enhance clinical treatment outcome. The results indicated that adults with untreated periodontitis harboring A. actinomycetemcomitans may benefit from the adjunctive antimicrobial therapy for a minimum of 12 months, whereas, the regimen may adversely affect the clinical treatment outcome of patients harboring P. gingivalis but not A. actinomycetemcomitans.

Adult↗

PubMed1998

Long-term maintenance of alveolar bone gain after implantation of autolyzed, antigen-extracted, allogenic bone in periodontal intraosseous defects.

This randomized controlled trial assessed the long-term maintenance of alveolar bone gain after implantation of autolyzed, antigen-extracted, allogenic (AAA) bone. AAA bone is a demineralized freeze-dried bone allograft processed after previously described methods. In each of 14 patients, AAA bone was implanted into the intraosseous defect of 1 tooth (test); a second tooth with an intraosseous defect was treated by modified Widman flap surgery alone (control). All patients were offered supportive periodontal therapy at 3- to 6-month intervals following treatment. Clinical measurements were taken prior to surgery, 6 months, and 3 years following surgery. Of the 14 patients enrolled, 11 patients completed the 6-month and 8 patients the 3-year examination. In test teeth, bone gain was significantly greater compared to control teeth at 6 months (2.2+/-0.5 mm and 1.2+/-0.5 mm, respectively) and 3 years (2.3+/-0.7 mm and 1.1+/-0.8 mm, respectively) (P < 0.05). Also, more probing attachment was gained in test compared to control teeth at 3 years (2.0+/-0.7 mm and 0.8+/-0.5 mm, respectively; P < 0.05). At 3 years, Porphyromonas gingivalis was detected in 3 test and 2 control teeth by polymerase chain reaction, whereas no Actinobacillus actinomycetemcomitans was found. Due to the low detection frequency, there was no clear correlation between the maintenance of alveolar bone during supportive periodontal therapy and subgingival infection with P. gingivalis. The data indicated that alveolar bone gain after implantation of AAA bone may be maintained over a minimum of 3 years in patients receiving periodontal supportive therapy.

Adult↗

PubMed1998

Working parameters of a magnetostrictive ultrasonic scaler influencing root substance removal in vitro.

This study assessed defect depth and volume resulting from root instrumentation using a magnetostrictive ultrasonic scaler with a slim scaling tip (P 12) in vitro. Combinations of the following working parameters were analyzed: lateral forces of 0.5 N, 1 N, and 2 N; tip angulations of 0 degrees, 45 degrees, and 90 degrees; power settings of low, medium, and high; and instrumentation time of 10, 20, 40, and 80 seconds. Defects were quantified using a 3-dimensional optical laser scanner. Lateral force had the greatest influence on defect volume compared to tip angulation and power setting (beta-weights 0.53+/-0.04, 0.17+/-0.04, and 0.19+/-0.04, respectively). Lateral force and tip angulation had similar effects on defect depth, but both effects were greater compared to power setting (beta-weights 0.43+/-0.04, 0.49+/-0.04, and 0.19+/-0.04, respectively). The various combinations of the assessed working parameters showed synergistic effects resulting in a wide range of defect depths (14.1+/-1.7 microm to 410.5+/-51.1 microm) and volumes (0.0084+/-0.0057 mm3 to 1.3+/-0.079 mm3). Severe root damage (defect depth >50 microm) at 40 seconds instrumentation time occurred under most combinations of lateral force, angulation, and power settings. The only exceptions were combinations of: 1) 0 degrees angulation, 0.5 N and 1 N lateral force at any power setting and 2) 45 degrees angulation, 0.5 N lateral force at low and medium power setting. The efficacy of the assessed magnetostrictive ultrasonic scaler may be adapted to the various clinical needs by adjusting the lateral force, tip angulation, and power setting.

Dental High-Speed Equipment↗

PubMed1997

Working parameters of a sonic scaler influencing root substance removal in vitro.

This study assessed defect depth and volume resulting from root instrumentation using a KaVo Sonic-flex Lux 2000 L sonic scaler with a slim scaling tip (Perio-Tip no. 8) in vitro. Combinations of the following working parameters were analyzed: lateral forces of 0.5 N, 1 N, and 2 N; tip angulations of 0 degree, 45 degrees, and 90 degrees; and instrumentation time of 10 s, 20 s, 40 s, and 80 s. Defects were quantified using a three-dimensional optical laser scanner. Instrumentation time had an almost linear impact on defect depth and volume. Although lateral force (beta-weight 0.55 +/- 0.062) had a greater influence on defect volume than tip angulation (beta-weight 0.29 +/- 0.062), their effects on defect depth were similar (beta-weight 0.43 +/- 0.052 and 0.50 +/- 0.052, respectively). The combination of force and angulation showed synergistic effects resulting in a wide range of defect depths (21.9 +/- 0.96 microns to 174 +/- 28.8 microns, at 40 s) and volumes (0.056 +/- 0.019 mm3 to 0.68 +/- 0.10 mm3 at 40 s). Severe root damage (> 50 microns/40 s) did not occur at any combination of 0.5 N lateral force and/or 0 degree tip angulation. By adjusting lateral force and tip angulation, the efficacy of the assessed sonic scaler may be adapted to various clinical needs.

Dental Instruments↗

PubMed1997

Controlled local delivery of tetracycline HCl in the treatment of periimplant mucosal hyperplasia and mucositis. A controlled case series.

The purpose of this controlled case series was to assess the adjunctive efficacy of controlled topical tetracycline HCl application in the treatment of infection associated periimplant mucositis or mucosal hyperplasia. Eight patients with at least 2 endosseous implants showing clinical signs of periimplant mucosal hyperplasia or mucositis were enrolled. All implants received supra- and subgingival scaling, with half of the implants receiving adjunctive controlled local delivery of tetracycline HCl (test). Control implants did not receive any other therapy aside from scaling. Clinical parameters were assessed at baseline, 4, and 12 weeks. Scaling plus controlled local delivery of tetracycline HCl markedly reduced periimplant mucosal hyperplasia in 4 of 5 test implants and demonstrated a trend towards a reduction of bleeding on probing scores. Scaling alone had no effect on mucosal hyperplasia in the 2 control implants presenting with this condition nor bleeding on probing scores. In both groups, plaque index scores were slightly reduced at 4 weeks but returned to baseline values at 12 weeks, whereas pocket probing depths, clinical attachment levels, and probing bone levels remained unchanged during the course of the trial. The observed trends suggest that scaling plus controlled local delivery of tetracycline HCl may have beneficial effects. Randomized controlled trials employing a sample size high enough to reach sufficient statistical power are needed to definitively assess the efficacy of controlled local tetracycline HCl delivery on periimplant diseases.

Administration, Topical↗

PubMed1996

Adjunctive controlled topical application of tetracycline HCl in the treatment of localized persistent or recurrent periodontitis. Effects on clinical parameters and elastase-alpha1-proteinase inhibitor in gingival crevicular fluid.

35 patients receiving regular supportive periodontal therapy (SPT) and showing signs of localized persistent or recurrent periodontitis were enrolled in the study. Within 1 week after SPT, each patient had a tetracycline HCl loaded ethylene vinyl acetate co-polymer fiber placed into the periodontal pocket of 1 randomly selected tooth with persistent or recurrent periodontitis (test); the fiber was removed after 9.5+/-2.0 days. A non-adjacent tooth with persistent or recurrent periodontitis in a separate quadrant, which received no further treatment, served as a control. A total of 28 patients completed the 6-month study. Compared to control teeth, in test teeth at 6 months significantly (p<0.01) lower scores were found for gingival index, pocket probing depths, and PMN elastase-alpha1-proteinase inhibitor concentrations in gingival crevicular fluid. With the exception of plaque index scores, test teeth demonstrated significant reductions from baseline to 6 months in all parameters (p<0.05). Conversely, all parameter measurements in control teeth, except bleeding on probing, showed no significant difference between baseline and 6-month values. The results suggest that the use of controlled topical application of tetracycline HCl may improve periodontal health and reduce the risk of disease progression in localized persistent or recurrent periodontitis. Moreover, the effects of this application appear to be sustained for at least 6 months.

Administration, Topical↗

PubMed1996

Efficacy of systemically administered acetylsalicylic acid plus scaling on periodontal health and elastase-alpha 1-proteinase inhibitor in gingival crevicular fluid.

The purpose of this proof of principle trial was to assess whether conventional periodontal therapy and systemically administrated acetylsalicylic acid (ASA) are functionally synergistic when combined in the treatment of periodontitis. A total of 30 patients with untreated moderate to severe adult periodontitis were enrolled into the study and were given placebo q.i.d. between the baseline and 6-week examination, and acetylsalicylic acid (ASA) 500 mg q.i.d. between the 6-week and 12-week examinations. In addition, they received supra- and subgingival scaling in 1 quadrant after baseline examination and in 2 further randomly selected quadrants after the 6-week examination. The study design resulted in the following 4 therapies: (1) scaling plus ASA 500 mg q.i.d.; (2) scaling plus placebo q.i.d.; (3) ASA 500 mg q.i.d. alone; (4) placebo q.i.d. alone. Two-way analysis of variance showed functional synergism of ASA and scaling, resulting in a therapeutic efficacy approximately equivalent to the sum of each individual therapeutic efficacy (i.e., ASA alone and scaling alone) in reducing gingival inflammation and pocket probing depth over the 6-week observation period (interaction: p > 0.05). Only the effect of ASA was significant in reducing the concentration of elastase-alpha 1-proteinase inhibitor in gingival crevicular fluid (GCF E-alpha 1-PI) (p < 0.001), reduction in GCF E-alpha 1-PI concentrations by ASA may indicate a decreased risk in periodontal disease progression. The results suggest that the combination of therapies and their different mechanisms of action, i.e., reduction of bacterial plaque and inhibition of destructive components of the immune responses, may result in functionally synergistic therapeutic efficacies in patients with untreated adult periodontitis.

Administration, Oral↗

PubMed1996

Serodiagnosis of Porphyromonas gingivalis infection by immunoblot analysis with recombinant collagenase.

The Porphyromonas gingivalis collagenase-specific serum immunoglobulin A (IgA), IgM, and IgG responses from 20 patients with early-onset periodontitis (EOP), 20 patients with adult periodontitis, (AP), and 20 age- and sex-matched healthy controls were examined by immunoblot analysis. A recombinant collagenase antigen used for the immunoblot analysis was produced by using the plasmid pGEX-2T, which allows the fusion between the collagenase and glutathione S-transferase. There was no significant difference in collagenase-specific IgG antibody detection between samples from the EOP, AP, and control groups. In contrast, 85% of AP and EOP sera had collagenase-specific IgA antibodies, whereas only 20% of control sera showed collagenase-specific IgA reactivity. Plaque samples from all groups were assessed by PCR with primers complementary to the collagenase-encoding gene prtC. The results indicated that 90% of AP and EOP plaque samples and 10% of control samples were positive for P. gingivalis. All patients with collagenase-specific IgA antibodies were PCR positive. The results of the study indicate a nearly complete concordance (k = 0.856) between the presence of collagenase-specific IgA antibodies and PCR detection of P. gingivalis. By using PCR as the "gold standard," the sensitivity and specificity of the IgA immunoblot test were 94.7 and 90.9%, respectively. Therefore, the recombinant collagenase is a potential candidate for use in the serodiagnosis of periodontitis.

Adult↗

PubMed1995

Adjunctive supragingival irrigation with acetylsalicylic acid in periodontal supportive therapy.

To assess the clinical efficacy of adjunctive supragingival irrigation with buffered 0.3% acetylsalicylic acid (ASA), 60 patients with periodontitis receiving supportive periodontal therapy were randomly assigned to 1 of 3 home regimens: (1) 1x daily adjunctive supragingival irrigation with 300 ml water immediately followed by 200 ml of buffered 0.3% ASA; (2) 1x daily adjunctive supragingival irrigation with 500 ml water; or (3) normal oral hygiene alone. Clinical parameters were assessed at baseline and 6 months. Irrigator use was measured by timers built into the irrigator units. Results at 6 months showed that both supragingival irrigation with buffered 0.3% ASA and supragingival irrigation with water significantly reduced gingival index scores (median 0.1 and 0.35, respectively) and pocket probing depths (both median 0.26 mm) compared to the control group. In addition, irrigation with water resulted in a significant reduction in bleeding on probing (median 0.13), whereas irrigation with buffered 0.3% ASA had no significant effect on bleeding on probing compared to the control group. The clinical efficacy of irrigation with either ASA or water was found to be positively correlated to initial disease severity and irrigator use. Thus, frequent supragingival irrigation with either 0.3% ASA or water in addition to regular oral hygiene appears to be a beneficial adjunct to periodontal supportive therapy in patients with moderate to severe signs of periodontitis. However, the use of buffered 0.3% ASA as an irrigant does not seem to enhance the clinical efficacy of supragingival irrigation on periodontal health.

Adult↗

PubMed1995

Identification of Actinobacillus actinomycetemcomitans in subgingival plaque by PCR.

The purpose of this study was to assess the sensitivity and specificity of the PCR in detecting Actinobacillus actinomycetemcomitans. The PCR's detection capability was compared with those of three other methods: culture-enhanced PCR (CE-PCR), colony hybridization (CH), and conventional culture with presumptive biochemical identification. A 285-bp stretch of the leukotoxin gene lktA of A. actinomycetemcomitans was amplified by PCR with primers TT-15 and TT-16. For CH, the PCR product was labeled with digoxigenin and used as a hybridization probe. Nucleotide sequence analysis of the PCR product of A. actinomycetemcomitans 1D4 and 1664 and three clinical isolates revealed complete homology among the tested strains, with only one base substitution (at position 1344) in comparison with the published sequence. With artificially infected subgingival plaque, the detection limit of PCR for A. actinomycetemcomitans was 10(3) CFU/ml of plaque suspension. Culturing subgingival plaque on tryptic soy-serum-bacitracin-vancomycin agar prior to PCR (CE-PCR) improved the limit of detection to 10(2) CFU/ml. Analysis of subgingival plaque samples from 35 patients with periodontal disease and 10 periodontally healthy subjects revealed that CE-PCR and CH had the highest overall rate of A. actinomycetemcomitans detection (both 58%), followed by PCR and culture (both 42%). With CH as the "gold standard", the sensitivities of CE-PCR, PCR, and culture were 88, 65, and 58%, respectively; the specificities were 84, 89, and 79%, respectively. The CE-PCR provided acceptable positive and negative predictive values (> or = 70%) when the prevalence of A. actinomycetemcomitans varied between 30 and 70%. PCR alone provided comparable predictive values over a narrower range of prevalence rates (30 to 50%), while culture did not afford acceptable predictive values at any prevalence rate. PCR and CE-PCR were found to be superior to culture with presumptive biochemical identification and should be the preferred methods for the detection of A. actinomycetemcomitans in subgingival plaque.

Aggregatibacter actinomycetemcomitans↗

PubMed1994

DNA fingerprinting of Eikenella corrodens by pulsed-field gel electrophoresis.

In order to conduct molecular typing of Eikenella corrodens strains by macrorestriction fingerprinting, we evaluated different restriction enzymes for digestion of genomic DNA and determined the optimal parameters for separating E. corrodens DNA by pulsed-field gel electrophoresis. Ten E. corrodens strains isolated from oral and extraoral infection sites in different individuals were analyzed. The rare-cutting restriction endonucleases DraI, SmaI and XbaI usually used for pulsed-field gel electrophoresis analyses were not suitable for digestion of E. corrodens genomic DNA because they either did not digest the DNA or produced bands of similar molecular weights that could not be separated. Accordingly, among additional enzymes including BamHI, BglII, EcoRI and Hind III, we found BamHI and BglII to be the most suitable rare-cutting enzymes for pulsed-field gel electrophoresis analysis. They cleaved the genomes of all the above strains into 15-20 fragment bands that were clearly separated by the following pulsed-field gel electrophoresis conditions: 140 V with a running time of 40 h, pulse times of 5 to 50 s with linear ramping and an electrical field angle of 120 degrees. These conditions enabled us to distinguish 8 individual pulsed-field gel electrophoresis patterns from the 10 strains analyzed. However, only 4 identical outer membrane protein profiles were differentiated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The data obtained in this analysis showed clonal divergence among members of the E. corrodens species, at the same time revealing this pulsed-field gel electrophoresis as being a highly attractive procedure for epidemiological investigation of this organism, including its acquisition and transmission.

Bacterial Outer Membrane Proteins↗

PubMed1994

Polymerase chain reaction for the identification of Porphyromonas gingivalis collagenase genes.

Porphyromonas gingivalis has been shown to exhibit genetic diversity possibly resulting in variation of virulence. In the present study a potential virulence factor was targeted for the detection of P. gingivalis. A 548 bp fragment of the collagenase gene (prtC) from Porphyromonas gingivalis ATCC 33277 was amplified by polymerase chain reaction (PCR) using oligonucleotides derived from the middle portion of prtC. From 16 of 21 clinical P. gingivalis strains, a PCR product of similar size to the prtC could be obtained. These 16 P. gingivalis strains were confirmed as positive for prtC using DNA hybridization with a digoxigenin-labeled prtC PCR product as a probe. In 12 of the 16 prtC positive strains, the restriction analysis of the PCR products revealed fragment patterns identical to the known sequence. In the other 4 prtC positive strains, 4 distinct patterns were found. Of these strains, nucleotide sequence analysis of a 400 bp PCR product stretch revealed 79.1%, 83.0%, 84.8 and 89.5% homology with the known nucleotide sequence for this specific region. Sequence analysis of the PCR products from the ATCC 33277 strain demonstrated 93.7% homology. The limit of detection for the PCR was about 100 organisms. None of the other 48 tested strains of 16 bacterial species derived from oral and extraoral infections yielded a PCR product. The PCR was also used for the detection of prtC sequences in dental plaque. Our data indicate that not all P. gingivalis strains have prtC. Nucleotide heterogeneity exists among P. gingivalis with prtC.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Proteins↗