Comparison of the detection of periodontal pathogens in bacteraemia after tooth brushing by culture and molecular techniques

Abstract

BackgroundThe prevalence and amounts of periodontal pathogens detected in bacteraemia samples after tooth brushing-induced by means of four diagnostic technique, three based on culture and one in a molecular-based technique, have been compared in this study.Material and MethodsBlood samples were collected from thirty-six subjects with different periodontal status (17 were healthy, 10 with gingivitis and 9 with periodontitis) at baseline and 2 minutes after tooth brushing. Each sample was analyzed by three culture-based methods [direct anaerobic culturing (DAC), hemo-culture (BACTEC), and lysis-centrifugation (LC)] and one molecular-based technique [quantitative polymerase chain reaction (qPCR)]. With culture any bacterial isolate was detected and quantified, while with qPCR only Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were detected and quantified. Descriptive analyses, ANOVA and Chi-squared tests, were performed.ResultsNeither BACTEC nor qPCR detected any type of bacteria in the blood samples. Only LC (2.7%) and DAC (8.3%) detected bacteraemia, although not in the same patients. Fusobacterium nucleatum was the most frequently detected bacterial species.ConclusionsThe disparity in the results when the same samples were analyzed with four different microbiological detection methods highlights the need for a proper validation of the methodology to detect periodontal pathogens in bacteraemia samples, mainly when the presence of periodontal pathogens in blood samples after tooth brushing was very seldom. Key words:Bacteraemia, periodontitis, culture, PCR, tooth brushing.

Comparison of two different methods for detecting periodontal pathogenic bacteria

“…However, this technique has some limitations, such as the difficulties to detect pathogens in sample containing small quantities of DNA, since more than 100 bacteria in periodontal pockets is required for PCR 9,10 . In addition, the exact amount of bacteria cannot be determined since it is a qualitative method of diagnostic, and therefore cannot be used to differentiate health and disease samples 6,8,9,11,12 . In order to overcome some of these drawbacks, different strategies of PCR have been introduced in many laboratories.…”

“…In order to overcome some of these drawbacks, different strategies of PCR have been introduced in many laboratories. More specifically, quantitative PCR (qPCR) allows the quantification of low number of bacterial loads in subgingival plaque samples [12][13][14] . qPCR is able to detect approximately 10 bacteria and is able to quantify the exact amount of bacteria present in subgingival crevicular fluid 6,12,15 .…”

Reduction in bacteremia after brushing with a triclosan/copolymer dentifrice—A randomized clinical study

“…Microbial culture, the standard for clinical diagnosis of bacteremic episodes, was utilized to determine blood borne bacteria (Jones, Munro, Grap, Kitten, & Edmond, 2010;Marín et al, 2016). This study did not directly analyse the type of bacteria recovered on agar plates.…”

Screening of Selected Plant-Derived Extracts for Their Antimicrobial Activity against Oral Pathogens

“…The most efficient way to prevent caries and other periodontal diseases is to remove the biofilm by brushing and flossing the teeth and conducting periodic dental cleaning or prophylaxis [5]. Unfortunately, most people fail to maintain a sufficient level of oral control through mechanical removal only, which has called for the use of oral products containing antimicrobial ingredients as a complementary measure to diminish biofilm formation on the tooth surface [6].…”