Rapid Quantification of Bacteria in Infected Root Canals Using Fluorescence Reagents and a Membrane Filter: A Pilot Study on Its Clinical Application to the Evaluation of the Outcomes of Endodontic Treatment

Abstract

Objective. The bacterial examination has been performed during the course of the root canal treatment. In the present pilot study, the new developed method, using fluorescence reagents and a membrane filter, was applied to the detection and quantification of bacteria in infected root canals, in order to evaluate the outcomes of the treatment. Methods. Six infected root canals with periapical lesions from 5 subjects were included. Informed consent was obtained from all subjects (age ranges, 23–79 years). Samples from infected root canals were collected at the beginning of the treatment (termed #25 First), the end of the first day of treatment (termed #55 First), and the next appointment day (termed #55 Second). Then, the bacterial count (CFU) was measured using fluorescence reagents (4′,6′-diamidino-2-phenylindole and propidium iodide) and the polycarbonate membrane filter by Bioplorer. Results. The mean ± SD of CFU in the sample of “#25 First” was (1.0 ± 1.4) × 105. As the root canal treatment progressed, the CFU decreased as 7.9 × 103 (#55 First) and 4.3 × 102 (#55 Second). Conclusion. In the present pilot study, rapid detection and quantification of bacteria in infected root canals were found to be successfully performed using fluorescence reagents and a membrane filter (Bioplorer analysis).

Rapid Bacterial Detection during Endodontic Treatment

“…Compared with more conventional microbiology techniques, such as CFU counting and polymerase chain reaction (Kato et al 2007;Sathorn et al 2007;Anderson et al 2012), the approach significantly reduces the processing time required, making it a viable technique for introduction into the current workstream of dental practices and capitalizing on otherwise discarded biological material. As opposed to recently published techniques (Sato et al 2012;Tan et al 2015), we demonstrate the detection of the residual bacteria directly on paper points, avoiding additional preparation of the sample or the root. Valuable time is saved, as measurements can be achieved in 5 min, making the technique clinically relevant for direct chair-side detection.…”

“…Furthermore, it remains unclear whether the sensitivity of such auto-fluorescence measurements would be sufficient to detect very low quantities of bacteria (Giana et al 2003;Sainsbury et al 2009;Ho et al 2010). Sato et al propose application of a system using live/dead staining and a membrane filter for bacterial counting (Sato et al 2012). Unfortunately, the procedure, despite being comparatively rapid, takes 30 minutes and requires additional collection and preparation of dentine samples, making it unfeasible for routine use in a clinical setting.…”

Is there bacterial infection in the intact crowns of teeth with pulp necrosis of sickle cell anaemia patients? A case series study nested in a cohort

“…Tenfold dilutions (up to 10 −3 ) of the lavage in phosphate‐buffered saline were transferred to the surface of plates containing blood agar for anaerobic microorganisms with 5% sheep blood (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), according to Sato et al . (2012). The plates were incubated under anaerobic condition at 37 °C for seven days.…”

Present status and future directions: Microbiology of endodontic infections

“…Enterococcus faecalis is one of the most prevalent species found in canals of teeth with post-treatment apical periodontitis (Gomes et al, 2008;Henriques et al, 2016;Murad et al, 2014;Peciuliene et al, 2000;Pinheiro et al, 2003Pinheiro et al, , 2015Rôças et al, 2004cSchirrmeister et al, 2007;Sedgley et al, 2006;Sundqvist et al, 1998). In terms of relative abundance, this species may account for a high range of less than 1% to 100% of the total bacteria Sanchez-Sanhueza et al, 2018;Sedgley et al, 2006;Zandi et al, 2016Zandi et al, , 2018.…”

“…Many of the areas of future research suggested above might serve as the basis for development of chairside tests to predict the treatment outcome or the risk of flare‐ups and other complications. Some chairside tests for rapid bacterial detection have been proposed in the literature (Herzog et al, 2017; Sato et al, 2012; Tan et al, 2015). However, they have yet to be introduced in the clinical routine, and there is a need for them to be validated by long‐term longitudinal follow‐up studies showing a correlation between the rapid chairside results and treatment outcome.…”