Comparison of real-time PCR and culture for detection of Porphyromonas gingivalis in subgingival plaque samples.
Porphyromonas gingivalis is a major pathogen in destructive periodontal disease in humans. Detection and quantification of this microorganism are relevant for diagnosis and treatment planning. The prevalence and quantity of P. gingivalis in subgingival plaque samples of periodontitis patients were determined by anaerobic culture and real-time PCR amplification of the 16S small-subunit rRNA gene. The PCR was performed with primers and a fluorescently labeled probe specific for the P. gingivalis 16S rRNA gene. By the real-time PCR assay, as few as 1 CFU of P. gingivalis could be detected. Subgingival plaque samples from 259 adult patients with severe periodontitis were analyzed. P. gingivalis was detected in 111 (43%) of the 259 subgingival plaque samples by culture and in 138 (53%) samples by PCR. The sensitivity, specificity, and positive and negative predictive values of the real-time PCR were 100, 94, 94, and 100%, respectively. We conclude that real-time PCR confirms the results of quantitative culture of P. gingivalis and offers significant advantages with respect to the rapidity and sensitivity of detection of P. gingivalis in subgingival plaque samples.
Boutaga
Khalil
K
Department of Periodontology, Section of Oral Microbiology, Academic Center for Dentistry Amsterdam, The Netherlands.
van Winkelhoff
Arie Jan
AJ
Vandenbroucke-Grauls
Christina M J E
CM
Savelkoul
Paul H M
PH
D016866:Bacteroidaceae Infections / Q000382:microbiology*; D015342:DNA Probes; D003773:Dental Plaque / Q000382:microbiology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D016966:Porphyromonas gingivalis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D015342:DNA Probes; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S