The additional value of real-time PCR in the quantitative detection of periodontal pathogens
: journal of clinical periodontology
Background and Aim: For the analysis of subgingival plaque, anaerobic bacterial
culture has been the gold standard for many years. Currently, molecular microbial
techniques have become available to identify and quantify target organisms with high
specificity and sensitivity. The technique of real-time (RT-PCR) provides a new tool to
detect oral pathogens both in oral and non-oral human infections. The aim of this study
was to compare the RT-PCR and anaerobic culture for detection and quantification of
six periodontal pathogens in periodontal health and disease.
Material and Methods: Subgingival plaque samples from 259 adult patients with
periodontitis and 111 healthy controls were analysed with quantitative anaerobic
culture and quantitative RT-PCR for Actinobacillus actinomycetemcomitans,
Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, Micromonas
micros and Fusobacterium spp.
Results: All species were more frequently isolated from patients than controls with
both culture and RT-PCR. P. gingivalis, T. forsythia and M. micros appeared
significant markers for disease with both techniques. P. intermedia was significantly
associated with periodontitis by RT-PCR only (OR 9.7), whereas A.
actinomycetemcomitans showed a significant relationship by culture only. The critical
differences between culture and RT-PCR were culture-negative/PCR-positive samples
which amounted to 7% for A. actinomycetemcomitans, 3% for P. gingivalis, 7% for
T. forsythia, 20% for P. intermedia, 6% for M. micros, and 0.8% for Fusobacterium
spp. in periodontitis patients and 12%, 3%, 2%, 35%, 14% and 0%, respectively, in the
periodontally healthy group. Furthermore, periodontitis individuals had significantly
higher amount of all of the test species in the subgingival plaque samples compared
with healthy subjects.
Conclusion: RT-PCR provides a new rapid diagnostic tool and opens the opportunity
to detect small numbers of oral pathogens in clinical specimens, which are under the
detection limit by culture technique.
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