Carpegen real-time polymerase chain reaction vs. anaerobic culture for periodontal pathogen identification.
OBJECTIVE
The aim of this study was to compare two methods of microbiological diagnosis, anaerobic bacterial culture and real-time polymerase chain reaction (PCR), for the detection of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, Fusobacterium nucleatum, and Treponema denticola.


METHODS
Seventy-two samples were collected from 18 patients who were suffering from aggressive periodontitis. The data obtained were compared for the two methods.


RESULTS
The results obtained with real-time PCR were different from those obtained with bacterial culture. The detection differences were 3% for A. actinomycetemcomitans, 8.33% for P. intermedia, and 12.5% for F. nucleatum. However, the differences for P. gingivalis and T. forsythia were 51.39% and 36.11%, respectively. No comparison was possible for T. denticola because it cannot be identified in culture. The variations found were the result of the better detection level (10(2) pathogens) of the PCR probe. Unlike bacterial culture, PCR allows the detection of T. denticola, which does not forming colonies and is oxygen sensitive. For F. nucleatum, T. forsythia and P. gingivalis, the real-time PCR technique was more sensitive than culture.


CONCLUSIONS
Good results were obtained with the real-time PCR technique for the six periopathogens targeted. This method seems to be indicated for its simplicity, rapidity and reproducibility but it cannot analyze data for an antibiotic susceptibility test. The periodontist must therefore choose one of these two methods according to his specific clinical objective: to obtain rapid, specific detection even with weak initial concentrations (but for targeted periopathogens only) or to be non-specific and analyze the pathological activity with an antibiogram.
Verner
C
C
Department of Periodontology, Faculty of Dentistry, Nantes, France.
Lemaitre
P
P
Daniel
A
A
Giumelli
B
B
Lakhssassi
N
N
Sixou
M
M
D016976:Aggregatibacter actinomycetemcomitans / Q000302:isolation & purification; D000693:Anaerobiosis; D015373:Bacterial Typing Techniques; D001431:Bacteriological Techniques; D001439:Bacteroides / Q000302:isolation & purification; D015169:Colony Count, Microbial / Q000379:methods; D004269:DNA, Bacterial / Q000032:analysis; D016967:Fusobacterium nucleatum / Q000302:isolation & purification; D006801:Humans; D010518:Periodontitis / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods; D016966:Porphyromonas gingivalis / Q000302:isolation & purification; D018713:Prevotella intermedia / Q000302:isolation & purification; D048373:Treponema denticola / Q000302:isolation & purification
D004269:DNA, Bacterial