Comparison of serological and DNA probe analyses for detection of suspected periodontal pathogens in subgingival plaque samples.
Several methods are currently being used to identify specific bacteria in dental plaque, namely direct culture, serological techniques and DNA probes. Culture methods are labour-intensive, dependent on the viability of the cells, and require fastidious growth conditions. Serological and DNA probes allow rapid strain-specific identification of periodontal pathogens with limited effort. The purpose of the present study was to compare results from serological and DNA probes in assessing and quantitating the presence of three suspected periodontal pathogens in subgingival plaque: Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bact. intermedius. Subgingival bacterial samples were obtained from 4 periodontal sites of each of 13 patients with untreated moderate to advanced adult periodontitis. Samples were taken using 2 paper points that were simultaneously inserted for 10 s to the bottom of the periodontal pockets. The plaque from one paper point was analysed by immunofluorescence using monoclonal antibodies. The other sample was analysed by species-specific cloned DNA probes. The number of bacteria per millilitre was calculated for both methods, and used for comparisons of results obtained with the two techniques. Results from indirect immunofluorescence and DNA hybridization analyses were negative for A. actinomycetemcomitans across all samples. Fluorescence did not detect bacteria at levels lower than 10(4), while the DNA probes identified organisms at levels of 10(3). Similar numbers of samples positive for Bact. gingivalis were obtained with either method (p = 0.227), and the results were not independent. A significantly greater proportion of Bact. intermedius-positive samples was detected by immunofluorescence (p = 0.0039), and the results of immunofluorescence and DNA hybridization were independent.(ABSTRACT TRUNCATED AT 250 WORDS)
Zappa
U
U
School of Dental Medicine, Department of Periodontology, University of Berne, Switzerland.
Reinking-Zappa
M
M
Graf
H
H
Gmür
R
R
Savitt
E
E
D000188:Actinobacillus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D000328:Adult; D000907:Antibodies, Bacterial / Q000032:analysis*; D000911:Antibodies, Monoclonal; D001439:Bacteroides / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis; D003773:Dental Plaque / Q000382:microbiology; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D008297:Male; D008875:Middle Aged; D010512:Periodontal Index; D010518:Periodontitis / Q000175:diagnosis / Q000382:microbiology*; D013045:Species Specificity
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D015342:DNA Probes; D004269:DNA, Bacterial