Comparison of culture and polymerase chain reaction techniques in the identification of Tannerella forsythia in periodontal health and disease, an in vitro study

Abstract

Background and Objectives:Various bacterial species from subgingival biofilm have demonstrated aetiological relevance in the initiation and progression of periodontitis. The aim of this study was to detect the presence of Tannerella forsythia (Tf) in subgingival plaque of periodontally healthy subjects and chronic periodontitis patients by using both culture and PCR technique and compare the two techniques.Materials and Methods:Pooled subgingival plaque samples were taken using sterile curettes from predetermined sites in 50 periodontally healthy subjects and from 50 periodontitis subjects. Samples were analyzed for the presence of T. forsythia using both techniques. Statistical analysis of the results was done using Chi-square test, sensitivity, and specificity tests.Results:Both techniques could detect T. forsythia in subgingival plaque samples from healthy and periodontitis subjects. Periodontally healthy individuals and individuals with chronic periodontitis using the culture technique showed the presence of T. forsythia in 14 and 34%, respectively. PCR technique showed the presence of T. forsythia in 20% healthy and 40% chronic periodontitis patients. T. forsythia detection in the periodontitis group was statistically significantly higher when compared to the healthy group by both culture and PCR technique (P = 0.019 and P = 0.029). PCR demonstrated high sensitivity and low specificity when compared to the culture technique.Conclusion:The results indicated that T. forsythia was more prevalent in periodontitis patients when compared with healthy subjects. The PCR was found to be more sensitive than culture technique for detection of T. forsythia from the subgingival plaque samples.

Periodontal Condition and Subgingival Microbiota Characterization in Subjects with Down Syndrome

“…These differences could be explained by the ability of qPCR to detect DNA of both viable and non-viable bacteria and to the lower detection limits of qPCR [32,38]. In addition, detection for T. forsythia by culturing is challenging [43,44], resulting in higher sensitivity and lower specificity when comparing qPCR technique versus culture [32,43,45]. However, the overall microbial tendency of an increase in prevalence for the three main target bacterial species from periodontal health to gingivitis and to periodontitis, was similar irrespective of the applied microbiological diagnostic technique.…”

Aggregatibacter actinomycetemcomitans

“…This would explain the fact that red complex bacteria can be present, although in very low numbers, in healthy individuals [121,124,125]. Moreover, a higher diversity and heterogeneity of the oral microbiome has been recognized recently.…”

“…It is important to note that even though defined microbial complexes are a convenient concept that has been commonly adopted worldwide, this view has been challenged by newer studies on the microbiome in periodontitis [119,120]. In fact, the increased use of advanced molecular-based technologies for the detection of bacteria, instead of the use of culture-based approaches only, has significantly advanced our understanding of the complexity of the periodontal microbiome in health and disease [119,[121][122][123]. For instance, the distinction between pathogens and commensals is not as clear as previously thought, as it is also dependent on the interactions of bacteria with the host.…”