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). MATERIALS AND 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. CONCLUSION: 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). CLINICAL RELEVANCE: 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.
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). MATERIALS AND 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. CONCLUSION: 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). CLINICAL RELEVANCE: 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: Juan Manuel Morillo; Laura Lau; Mariano Sanz; David Herrera; Conchita Martín; Augusto Silva Journal: J Clin Periodontol Date: 2004-12 Impact factor: 8.728
Authors: Ellen Teresa Boehm; Cosima Thon; Juozas Kupcinskas; Ruta Steponaitiene; Jurgita Skieceviciene; Ali Canbay; Peter Malfertheiner; Alexander Link Journal: Sci Rep Date: 2020-10-01 Impact factor: 4.379
Authors: Pune N Paqué; Christopher Herz; Joël S Jenzer; Daniel B Wiedemeier; Thomas Attin; Nagihan Bostanci; Georgios N Belibasakis; Kai Bao; Philipp Körner; Tanja Fritz; Julia Prinz; Patrick R Schmidlin; Thomas Thurnheer; Florian J Wegehaupt; Konstantinos Mitsakakis; Johannes R Peham Journal: J Clin Med Date: 2020-09-11 Impact factor: 4.241