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Int Poster J Dent Oral Med 9 (2007), No. 3 15. Sep. 2007
Int Poster J Dent Oral Med 2007, Vol 9 No 03, Poster 374
Two Subgingival Plaque Sampling Strategies Used With RNA-Probes
Language: English
Authors:
Dr. med. dent. Diana Krigar, Dr. med. dent. Jens Kaltschmitt, Dr. med. dent. Jörg K. Krieger
Section of Periodontology, Department of Conservative Dentistry, Clinic for Oral, Dental and Maxillofacial Diseases, University Hospital Heidelberg
Prof. Dr. med. dent. Peter Eickholz
Department of Periodontology, Center for Dental, Oral, and Maxillofacial Medicine , University Hospital Frankfurt
Date/Event/Venue:
29, 30 June and 1 July 2006
Europerio 5
Madrid, Spain
Introduction
Of a total of about 500 bacterial species colonizing the oral cavity, some
are narrowly assoziated with periodontal destruction [Moore & Moore 1994].
Actinobacillus actinomycetemcomitans, Tannerella forsythensis, Porphyromonas
gingivalis, and Treponema denticola belong to the periodontal pathogens
[Socransky et al. 1998]. It has been shown that Actinobacillus
actinomycetemcomitans (AA) has an important role in the etiology of
aggressive periodontal disease [Bragd et al. 1985, Newman et al. 1976]. AA
is a microaerophilic, facultative anaerobic, and Gram-negative coccoid rod
belonging to the family of Pasteurellaceae. Periodontal disease associated
with AA in many cases cannot be treated reliably and predictively by
mechanical removal of the subgingival biofilm alone [Christersson et al.
1985, Kornman et al. 1985, Mombelli et al. 1994]. Thus, the detection of AA
is a significant factor contributing to the decision whether mechanical
antiinfective therapy should be adjuncted by systemic antibiotics. Depending
on the microbial complexes that are detected from subgingival plaque
different antibiotic regimes are recommended [Beikler et al. 2004].
Microbiological testing prior to antiinfective therapy is recommended for
the following clinical diagnoses: aggressive periodontitis, generalised
severe chronic periodontitis, refractory periodontitis, and severe
periodontitis associated with systemic diseases (e.g. HIV infection)
[Flemmig et al. 1998]. Subgingival plaque samples should be taken from the
deepest pockets exhibiting signs of activity, i.e. bleeding or suppuration.
A micribiological analysis representative for the subgingival microflora of
the whole oral cavity is relevant for adjunctive systemic antibiotic therapy
of certain forms of periodontitis. Therefore also due to commercial reasons
the analysis of pooled plaque sampled from several sites is recommended
[Flemmig et al. 1998]. Thus, in this study the results of separate
microbiological RNA-probe analyses of subgingival plaque samples from 3
different sites should be compared with the results from a pooled
sample.
Material and Methods
 |
 |
Abb.1 |
Patients
- 158 patients (72 female), 47.4 ± 10.6 years of age
- Two indications for microbiological examination:
- diagnosis of untreated aggressive, generalised severe chronic periodontitis or periodontitis as manifestation of systemic disease
- reevaluation after combined non-surgically mechanical and systemic antibiotic treatment of A.a.-associated periodontitis (after antibiotic therapy)
Clinical examinations
- At 6 sites per tooth PD and CAL-V using a rigid periodontal probe (PCPUNC 15, Hu Friedy, Chicago IL, USA)
- reference for CAL-V measurement CEJ or margin of restoration, BOP 30 sec. after probing
Microbiological examination
- 3 deepest pockets in 3 different quadrants; after removing the supragingival plaque, drying the test site by air and held dry using cotton rolls
- Simultaneous insertion of 2 sterile paper points to the bottom of the pocket and removal after 10 seconds
- One paper point of each site was put into a separate transportation vial, the other was pooled with paper points of the respective 2 other sampling sites (MT3)
- Analysis: RNA probe test kit (IAI Pado Test 4.5®, Institut für angewandte Immunologie, Zuchwil, Schweiz) with a detection limit of 103,3 for Actinobacillus actinomycetemcomitans (AA), Porphyromonas gingivalis (PG), Tannerella forsythensis (TF), Treponema denticola (TD)
- 76 patients received antiinfective therapy (oral hygiene instructions, professional tooth cleaning, supra- and subgingival scaling at all teeth within 24 hours according to the concept of "full-mouth-disinfection")
- In all 76 patients AA had been detected subgingivally before therapy and, thus mechanical therapy was combined with the systemic administration of 375mg and 250mg metronidazole 3 times daily/7days.
Statistical analysis
- All bacterial counts underwent logarithmic transformation
- Two variables were analysed for each periodontal pathogen:
- log-transformed bacterial counts prevalence, i.e. detection of the pathogen or not
- Prevalence of a microorganism was assessed if at least in one of the 3 samples the respective pathogen had been detected.
- Prevalence for separate analysis and MT3 were compared using Wilcoxon signed ranks tests for paired samples
- Statistical analyses were done using SystatTM for Windows Version 10, Systat Inc. Evanston, USA
Results
- The mean log-transformed number of bacteria was higher in pooled samples
than the mean value of the results of the separate samples of all tested
pathogens (p<0.001).
- However, for all 4 pathogens analysis failed to detect
statistically significant differences between the separate samples and the
mt3 regarding the detection frequency. These findings were observed over all
samples as well as after evaluation of samples before and after
separately.
|
PD |
CAL-V |
All (n=158) |
7.17 ± 1.9 |
8.00 ± 2.00 |
Before therapy (n=82) |
8.07 ± 1.49 |
8.45 ± 1.59 |
After therapy (n=76) |
6.19 ± 1.81 |
7.43 ± 2.23 |
ρ |
<0.001 |
<0.001 |
Tab.1 Clinical parameters |
|
Seperate Analysis |
MT3 |
P |
AA |
1.01 ± 1.43 |
1.92 ± 2.37 |
<0.001 |
TF |
4.02 ± 2.41 |
5.28 ± 2.38 |
<0.001 |
PG |
4.04 ± 2.46 |
5.20 ± 2.48 |
<0.001 |
TD |
3.88 ± 2.30 |
4.81 ± 2.47 |
<0.001 |
Tab.2 Log-transformed Bacterial Counts for Separate and Pooled Analysis (MT3) |
AA |
|
Seperate analysis |
|
|
Not detected |
detected |
Total |
MT3 |
Not detected |
68 |
23 |
96 |
|
detected |
26 |
41 |
67 |
Total |
|
94 |
64 |
158 |
Tab.3 Prevalence of AA, TF, PG,TD
for Separate and Pooled Analysis (MT3) No statistically significant
difference between separate analysis and MT3 detected (p=0.668) |
TF |
|
Seperate analysis |
|
|
Not detected |
detected |
Total |
MT3 |
Not detected |
15 |
11 |
26 |
|
detected |
10 |
122 |
132 |
Total |
|
25 |
133 |
158 |
Tab.3 Prevalence of AA, TF, PG,TD
for Separate and Pooled Analysis (MT3) No statistically significant
difference between separate analysis and MT3 detected (p=0.827) |
PG |
|
Seperate analysis |
|
|
Not detected |
detected |
Total |
MT3 |
Not detected |
14 |
10 |
24 |
|
detected |
13 |
121 |
134 |
Total |
|
27 |
131 |
158 |
Tab.3 Prevalence of AA, TF, PG,TD
for Separate and Pooled Analysis (MT3) No statistically significant
difference between separate analysis and MT3 detected (p=0.505) |
TD |
|
Seperate analysis |
|
|
Not detected |
detected |
Total |
MT3 |
Not detected |
14 |
10 |
24 |
|
detected |
17 |
117 |
134 |
Total |
|
31 |
127 |
158 |
Tab.3 Prevalence of AA, TF, PG,TD
for Separate and Pooled Analysis (MT3) No statistically significant
difference between separate analysis and MT3 detected (p=0.178) |
Conclusions
Pooling of subgingival plaque samples increased the bacterial counts per
analysis compared to separate samples and thus may increase the probability
to detect existing pathogens. However, this observation had no statistically
significant effect on the detection frequency of the tested pathogens.
Literature
- Armitage, G. C.: Development of a classification system for periodontal diseases and conditions. Ann Periodontol 4, 1-6 (1999)
- Beikler, T., Prior, K., Ehmke, B., Flemmig T. F.: Specific antibiotics in the treatment of periodontitis - A proposed strategy. J Periodontol 75, 169-175 (2004)
- Bragd, L., Dahlen, G., Wikström, M., Slots, J.: The capacity of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius to indicate progressive periodontitis; retrospective study. J Clin Periodontol 14, 95 (1985).
- Christersson, L., Fransson, C., Dunford, R., Zambon, J.: Microbiological and clinical effects of surgical treatment of localized juvenile periodontitis. J Peridontol 63, 418 (1992).
- Flemmig, T. F., Christgau, M., Karch, H.: Mikrobiologische Diagnostik bei marginalen Parodontopathien. Gemeinsame Stellungnahme der Deutschen Gesellschaft für Parodontologie (DGP) und der Deutschen Gesellschaft für Zahn-, Mund- und Kieferheilkunde (DGZMK). Dtsch Zahnärztl Z 53, 825-826 (1998).
- Kornman, K., Robertson, P.: Clinical and microbiological evaluation of therapy of juvenile peridontitis. J Periodontol 56, 443 (1985).
- Moore, W. E. C., Moore, L. V. H.: The bacteria of periodontal diseases. Periodontol 2000 5, 66-77 (1994).
- Mombelli, A., Gmür, R., Gobbi, C., Lang, N. P.: Actinobacillus actinomycetemcomitans in adult periodontitis. I. Topographic distribution before and after treatment. J Periodontol 65, 820-826 (1994).
- Quirynen, M., Bollen, C. M. L., Vandekerckhove, B. N. A., Dekeyser, C., Papaioanou, W., Eyssen, H.: Full- vs. Partial-mouth disinfection in the treatment of periodontal infections: short-term clinical and microbiological observations. J Dent Res 74, 1459 (1995).
- Socransky, S. S., Haffajee, A. D., Cugini, M. A., Smith, C., Kent Jr., R. L.: Microbial complexes in subgingival plaque. J Clin Periodontol 25, 134-144 (1998).
This Poster was submitted by Dr. med. dent. Diana Krigar.
Correspondence address:
Dr. med. dent. Diana Krigar
Section of Periodontology, Department of Conservative Dentistry, Clinic for Oral, Dental and Maxillofacial Diseases
University Hospital Heidelberg
Im Neuenheimer Feld 400
D-69120 Heidelberg
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