Rapid detection and quantification of five periodontopathic bacteria by real-time PCR.
The quantity of periodontopathic bacteria in plaque samples is an important determinant for understanding the etiologic role of bacteria. The real-time PCR method was used to detect and quantify periodontopathic bacteria, such as Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Porphyromonas gingivalis, Treponema denticola, and Treponema socranskii, in saliva and subgingival plaque samples. There was good agreement between the results of conventional PCR and real-time PCR methods. Using the LightCycler system we were able to determine the amount of periodontopathic bacteria within an hour. The real-time PCR method was linear for samples containing from 10(3) to more than 10(8) cells (r2 = 0.999). The application of the real-time PCR method should be useful in the rapid detection and quantification of periodontopathic bacteria in clinical samples.
Japan Collection of Microorganisms, RIKEN, Wako, Saitama. sakaomto@jcm.riken.go.jp
M
M
Sakamoto
Y
Y
Takeuchi
M
M
Umeda
I
I
Ishikawa
Y
Y
Benno
D000188:Actinobacillus / Q000254:growth & development / Q000302:isolation & purification; D000328:Adult; D001431:Bacteriological Techniques; D001439:Bacteroides / Q000254:growth & development / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000032:analysis; D006801:Humans; D008875:Middle Aged; D010510:Periodontal Diseases / Q000382:microbiology; D010519:Periodontium / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D016966:Porphyromonas gingivalis / Q000254:growth & development / Q000302:isolation & purification; D012463:Saliva / Q000382:microbiology*; D012680:Sensitivity and Specificity; D014208:Treponema / Q000254:growth & development / Q000302:isolation & purification
D004269:DNA, Bacterial