Comparison of culture methods and multiplex PCR for the detection of periodontopathogenic bacteria in biofilm associated with severe forms of periodontitis.
Conventional culture methods and Multiplex PCR, both of which we have been used for a long time in our clinical microbiology laboratory, were compared for their ability to detect a selected panel of periodontopathic bacteria: Aggregatibacter actinomycetemcomitans, Campylobacter rectus, Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas gingivalis, and Prevotella intermedia. Tests were performed in a single subgingival sample taken from a periodontal diseased site with a probing depth equal to or greater than 6mm. The results were compared site-by-site, taking into account the quality and the presence or absence of pathogens. 529 samples of subgingival plaque were analysed and the prevalence of the six species monitored varied in relation to the species itself and the method of detection. The most represented species is F. nucleatum, with a percentage of positive variability between 44.9% PCR and 46.5% culture test. Generally, the lowest prevalence was determined by culture test, with the exception of E. corrodens and F. nucleatum, which, unlike other bacteria, have been seen in higher percentages in culture with respect to PCR. For both methods, there was a good degree of accuracy in the determination of A. actinomycetemcomitans, C. rectus, E. corrodens, and P. gingivalis. It becomes weak for F. nucleatum and P. intermedia. Both culture and PCR techniques introduced many methodological problems when applied in oral microbiology, but the ideal technique for accurate detection of pathogens in subgingival plaque samples has yet to be developed.
Oral Microbiology Laboratory, Department of Biomedical Sciences, School of Medicine, "G. d'Annnunzio" University of Chieti-Pescara, Italy.
Simonetta
S
D'Ercole
Giovanni
G
Catamo
Domenico
D
Tripodi
Raffaele
R
Piccolomini
D001431:Bacteriological Techniques; D018441:Biofilms; D003773:Dental Plaque / Q000382:microbiology; D006090:Gram-Negative Bacteria / Q000302:isolation & purification* / Q000502:physiology; D006801:Humans; D025202:Molecular Diagnostic Techniques; D010518:Periodontitis / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods; D015995:Prevalence; D012680:Sensitivity and Specificity