Comparison of subgingival bacterial sampling with oral lavage for detection and quantification of periodontal pathogens by real-time polymerase chain reaction.
BACKGROUND
Saliva has been studied for the presence of subgingival pathogens in periodontitis patients. With the anaerobic culture technique, the discrepancy between salivary recovery and subgingival presence has been significant, which makes this approach not suitable for practical use in the microbial diagnosis of periodontitis patients. The real-time polymerase chain reaction (PCR) technique represents a very sensitive technique to detect and quantify bacterial pathogens. The aim of the study was to compare the presence and numbers of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythensis, Prevotella intermedia, and Micromonas micros in subgingival plaque and mouthwash samples by the anaerobic culture and real-time PCR techniques.


METHODS
Pooled subgingival plaque samples and 10-ml mouthwash samples were collected from 21 adult patients with periodontitis and analyzed by quantitative anaerobic culture and real-time PCR for A. actinomycetemcomitans, P. gingivalis, T. forsythensis, P. intermedia, and M. micros.


RESULTS
The detection frequency of A. actinomycetemcomitans, P. gingivalis, and T. forsythensis in subgingival plaque was identical by culture and real-time PCR and was higher for P. intermedia and M. micros by real-time PCR. The highest detection frequencies for the target bacteria were found in mouthwash samples by real-time PCR. The additional value of the real-time PCR to detect target bacteria was 38% for P. gingivalis, 73% for T. forsythensis, 77% for P. intermedia, and 71% for M. micros. The sensitivity to detect target species in mouthwash by real-time PCR was 100% for all test species except for P. intermedia (93.8%).


CONCLUSIONS
Rapid detection and quantification of periodontal pathogens in mouthwash samples are possible by real-time PCR. The procedure is significantly less time-consuming than subgingival sampling with paper points. This approach to detect major periodontal pathogens in mouthwash samples may simplify microbial diagnosis in periodontitis patients and may be used to monitor periodontal treatment.
Boutaga
Khalil
K
Department of Oral Microbiology, Academic Center for Dentistry Amsterdam, University of Amsterdam and Vrije University, Amsterdam, The Netherlands.
Savelkoul
Paul H M
PH
Winkel
Edwin G
EG
van Winkelhoff
Arie J
AJ
D000328:Adult; D016976:Aggregatibacter actinomycetemcomitans / Q000235:genetics / Q000302:isolation & purification; D001421:Bacteria, Anaerobic / Q000235:genetics / Q000302:isolation & purification*; D001439:Bacteroides / Q000235:genetics / Q000302:isolation & purification; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000032:analysis; D003773:Dental Plaque / Q000382:microbiology*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009067:Mouthwashes; D010462:Peptostreptococcus / Q000235:genetics / Q000302:isolation & purification; D010514:Periodontal Pocket / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods; D016966:Porphyromonas gingivalis / Q000235:genetics / Q000302:isolation & purification; D018713:Prevotella intermedia / Q000235:genetics / Q000302:isolation & purification; D012463:Saliva / Q000382:microbiology*; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial; D009067:Mouthwashes