Quantitative detection of volatile sulfur compound- producing microorganisms in oral specimens using real-time PCR.
OBJECTIVE
It is well-known that some periodontopathic bacteria, especially Porphyromonas gingivalis, Fusobacterium nucleatum, Tannerella forsythia (formerly Bacteroides forsythus or Tan. forsythensis), and Treponema denticola, actively produce volatile sulfur compounds (VSCs), such as H2S and CH3SH. We previously reported a qualitative relationship between periodontopathic bacteria and VSCs; however, a quantitative analysis of periodontopathic bacteria in oral specimens is required for further characterization of the relationship between oral bacteria and VSCs. In this study, we report a real-time polymerase chain reaction (PCR) method for the quantitative analysis of VSC-producing bacteria in oral specimens.


METHODS
Specimens were collected from 22 patients who visited the Preventive Dentistry and Breath Odor Clinic of Kyushu Dental College. A real-time PCR assay using the TaqMan system, based on the 5'-3' exonuclease activity of Taq polymerase, was employed for the quantitative analysis of periodontopathic bacteria that produce VSCs.


RESULTS
Using real-time PCR, we performed a quantitative analysis of P. gingivalis, F. nucleatum, Tan. forsythia, and T. denticola in the saliva, on the tongue coat, and in the subgingival plaque of patients with oral malodor.


CONCLUSIONS
Real-time PCR using the TaqMan system can be used for the quantitative analysis of VSC-producing oral bacteria.
Kato
H
H
Department of Preventive Dentistry, Kyushu Dental College, Kitakyushu, Japan. akihiro@kyu-dent.ac.jp
Yoshida
A
A
Awano
S
S
Ansai
T
T
Takehara
T
T
D000328:Adult; D000368:Aged; D015373:Bacterial Typing Techniques; D001439:Bacteroides / Q000302:isolation & purification / Q000378:metabolism; D004269:DNA, Bacterial / Q000032:analysis; D003773:Dental Plaque / Q000382:microbiology; D005260:Female; D016967:Fusobacterium nucleatum / Q000302:isolation & purification / Q000378:metabolism; D006209:Halitosis / Q000382:microbiology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D016966:Porphyromonas gingivalis / Q000302:isolation & purification / Q000378:metabolism; D012463:Saliva / Q000382:microbiology; D013457:Sulfur Compounds / Q000378:metabolism; D019914:Taq Polymerase; D014059:Tongue / Q000382:microbiology; D048373:Treponema denticola / Q000302:isolation & purification / Q000378:metabolism
D004269:DNA, Bacterial; D013457:Sulfur Compounds; D019914:Taq Polymerase