Comparing culture, real‐time PCR and fluorescence resonance energy transfer technology for detection of Porphyromonas gingivalis in patients with or without peri‐implant infections

Abstract

Galassi F, Kaman WE, Anssari Moin D, van der Horst J, Wismeijer D, Crielaard W, Laine ML, Veerman ECI, Bikker FJ, Loos BG. Comparing culture, real‐time PCR and fluorescence resonance energy transfer technology for detection of Porphyromonas gingivalis in patients with or without peri‐implant infections. J Periodont Res 2012; 47: 616–625. © 2012 John Wiley & Sons A/S Background and Objective: The aim of the study was to compare the detection of Porphyromonas gingivalis using a fluorescence resonance energy transfer (FRET) technology with commonly used diagnostic methods in salivary and subgingival plaque samples from subjects with dental implants. P. gingivalis was considered as a marker for a pathogenic microbiota. Material and Methods: Ninety‐seven adult subjects were recruited, including periodontally healthy controls with no dental implants, implant controls with no peri‐implant disease and patients with peri‐implant disease. Saliva and subgingival/submucosal plaque samples were collected from all subjects and were analyzed using culture, real‐time PCR and FRET technology employing P. gingivalis‐specific substrates. Results: It was found that the P. gingivalis‐specific substrates were highly suitable for detecting the presence of P. gingivalis in saliva and in subgingival plaque samples, showing comparable specificity to culture and real‐time PCR. Conclusion: We applied the FRET technology to detect P. gingivalis in implant patients with or without an implant condition and in controls without implants. The technique seems suitable for detection of P. gingivalis in both plaque and saliva samples. However, with all three techniques, P. gingivalis was not very specific for peri‐implantitis cases. Future work includes fine‐tuning the FRET technology and also includes the development of a chair‐side application.

Microbiome of peri-implant infections: Lessons from conventional, molecular and metagenomic analyses

“…PCR analysis offers great sensitivity as it allows a small amount of DNA of the microbial sample to be amplified many times in an exponential manner. It has been used for the analysis of microbial samples from dental implants for the detection of A. actinomycetemcomitans, 30 Porphyromonas gingivalis, 30,31 Prevotella intermedia, 30,32 Fusobacterium spp. 32 The DNA-DNA hybridization ('checkerboard') technique belongs to molecular techniques, developed by Socransky and co-workers.…”

Learned and unlearned concepts in periodontal diagnostics: a 50‐year perspective

“…Under the appropriate conditions these bacteria can be important contributors to the initiation and progression of periodontal infections. However, virtually all of the putative pathogens can also be found at low levels in many periodontally healthy people (15,16,52,56,60,61,65,82,83,103,120,126,135,159,163,181,195,196,216,263,266,267,281,288,291,292,299) and at healthy peri-implant sites (38,40,72,129,276). One interpretation of this observation is that these bacteria are a normal part of the health-associated oral microbiota and only become pathogens when there is a major disturbance in the host-microbe homeostasis.…”

Antibiotic resistance in human peri‐implantitis microbiota

“…These species have also been implicated as putative pathogens in the pathogenesis of periodontitis (Slots et al 1988;Rams et al 1990Rams et al , 1992Socransky et al 1998). Additional bacterial species and novel phylotypes may be identified in peri-implantitis lesions with other types of selective culture media (Rams et al 1990(Rams et al , 1992 and with various nucleic acid-based molecular methods, such as checkerboard DNA-DNA hybridization (Leonhardt et al 2003a), end-point and real-time PCR (Sato et al 2011;Galassi et al 2012), denaturing gradient gel electrophoresis (Zijnge et al 2006;Al-Radha et al 2012), and 16S ribosomal RNA sequencing (Koyanagi et al 2010;Kumar et al 2012). It is not known from the present retrospective study design whether these organisms initiated periimplantitis similar to periodontitis, or merely colonized deep peri-implant probing depths secondarily after other etiologic factors triggered the onset of peri-implant marginal tissue breakdown.…”