Estimation of total bacteria by real-time PCR in patients with periodontal disease.
BACKGROUND
Periodontal diseases are associated with the presence of elevated levels of bacteria within the gingival crevice.


OBJECTIVE
The aim of this study was to evaluate a total amount of bacteria in subgingival plaque samples in patients with a periodontal disease.


METHODS
A quantitative evaluation of total bacteria amount using quantitative real-time polymerase chain reaction (qRT-PCR) was performed on 20 samples of patients with ulceronecrotic periodontitis and on 10 samples of healthy subjects. The estimation of total bacterial amount was based on gene copy number for 16S rRNA that was determined by comparing to Ct values/gene copy number of the standard curve.


RESULTS
A statistically significant difference between average gene copy number of total bacteria in periodontal patients (2.55 x 10⁷) and healthy control (2.37 x 10⁶) was found (p = 0.01). Also, a trend of higher numbers of the gene copy in deeper periodontal lesions (> 7 mm) was confirmed by a positive value of coefficient of correlation (r = 0.073).


CONCLUSIONS
The quantitative estimation of total bacteria based on gene copy number could be an important additional tool in diagnosing periodontitis.
Gavrilo
G
Brajović
Branka
B
Popović
Miljan
M
Puletić
Marija
M
Kostić
Jelena
J
Milasin
D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification; D058491:Bacterial Load / Q000379:methods*; D016022:Case-Control Studies; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D006801:Humans; D010510:Periodontal Diseases / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
D004269:DNA, Bacterial