title: Real-time PCR quantification of six periodontal pathogens in saliva samples from healthy young adults.
abstract: OBJECTIVE
  The use of saliva as a diagnostic fluid for the evaluation of periodontal health has gained attention recently. Most published real-time PCR assays focused on quantification of bacteria in subgingival plaque, not in saliva. The aims of this study were to develop a real-time PCR assay for quantification of six periodontal pathogens in saliva and to establish a relationship between the amount of DNA (fg) and colony-forming unit (CFU).
  
  
  METHODS
  TaqMan primers/probe sets were used for the detection of Aggregatibacter actinomycetemcomitans (Aa), Eikenella corrodens (Ec), Fusobacterium nucleatum (Fn), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Tannerella forsythia (Tf), and total bacteria. Six periodontal pathogens and total bacteria in saliva from 24 periodontally healthy individuals were determined. The relationship between the amount of DNA (fg) and CFU was established by measuring the concentrations of extracted bacterial DNA and CFU per milliliter of bacteria on agar plates.
  
  
  RESULTS
  Fn, Ec, and Pi were detected in all saliva samples, while 58.5, 45.8, and 33.3% were detected for Tf, Pg, and Aa, respectively. Numbers of Ec and Fn in saliva were highly correlated (R(2) = 0.93, P < 0.01). The values of DNA (fg) per CFU ranged from 64 for Ec to 121 for Pg.
  
  
  CONCLUSIONS
  The real-time PCR assay in combination with the relationship between DNA (fg) and CFU can be used to quantitate periodontal pathogens in saliva and estimate the number of live bacteria (CFU).
  
  
  CONCLUSIONS
  This real-time PCR assay in combination with the relationship between DNA (fg) and CFU has the potential to be an adjunct in evaluation of periodontal health status.
authors.affiliation: Center for Dental Research, Loma Linda University School of Dentistry, 24876 Taylor Street, Loma Linda, CA, 92350, USA, zhou121@hotmail.com.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Zhou
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Liu
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Raydolfo M
authors.initials: RM
authors.lastname: Aprecio
authors.forename: Wu
authors.initials: W
authors.lastname: Zhang
authors.forename: Yiming
authors.initials: Y
authors.lastname: Li
mesh_terms: D000328:Adult; D001419:Bacteria / Q000235:genetics; D058491:Bacterial Load / Q000379:methods*; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D006801:Humans; D010518:Periodontitis / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012463:Saliva / Q000382:microbiology*; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D004269:DNA, Bacterial