Discrepancy between culture and DNA probe analysis for the detection of periodontal bacteria.
The purpose of this study was to compare a commercially available DNA probe technique with conventional cultural techniques for the detection of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in subgingival plaque samples. Samples from 20 patients with moderate to severe periodontitis were evaluated at baseline and during a 15 months period of periodontal treatment. Paperpoints from 4 periodontal pockets per patient were forwarded to Omnigene for DNA probe analysis, and simultaneously inserted paperpoints from the same pockets were analyzed by standard culture techniques. In addition, mixed bacterial samples were constructed harbouring known proportions of 25 strains of A. actinomycetemcomitans, P. gingivalis and P. intermedia each. A relatively low concordance was found between both methods. At baseline a higher detection frequency was found for A. actinomycetemcomitans and P. gingivalis for the DNA probe technique; for P. intermedia the detection frequency by culture was higher. For A. actinomycetemcomitans, 21% of the culture positive samples was positive with the DNA probe. Testing the constructed bacterial samples with the DNA probe method resulted in about 16% false positive results for the 3 species tested. Furthermore, 40% of P. gingivalis strains were not detected by the DNA probe. The present data suggest that at least part of the discrepancies found between the DNA probe technique used and cultural methods are caused by false positive and false negative DNA probe results. Therefore, the value of this DNA probe method for the detection of periodontal pathogens is questionable.
Department of Oral Microbiology, Academic Centre for Dentistry, Amsterdam, The Netherlands.
T J
TJ
van Steenbergen
M F
MF
Timmerman
F H
FH
Mikx
G
G
de Quincey
G A
GA
van der Weijden
U
U
van der Velden
J
J
de Graaff
D000328:Adult; D016976:Aggregatibacter actinomycetemcomitans / Q000235:genetics / Q000302:isolation & purification; D001431:Bacteriological Techniques; D003470:Culture Media; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis; D003773:Dental Plaque / Q000382:microbiology; D005188:False Negative Reactions; D005189:False Positive Reactions; D005260:Female; D005500:Follow-Up Studies; D006090:Gram-Negative Bacteria / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008875:Middle Aged; D010514:Periodontal Pocket / Q000382:microbiology; D010518:Periodontitis / Q000382:microbiology* / Q000628:therapy; D016966:Porphyromonas gingivalis / Q000235:genetics / Q000302:isolation & purification; D018713:Prevotella intermedia / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity
D003470:Culture Media; D015342:DNA Probes; D004269:DNA, Bacterial