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      Development of a point-of-care-device for fast detection of periodontal pathogens

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          Abstract

          Background

          A number of pathogens can cause severe destruction of the periodontal apparatus during the course of periodontitis. The aim of this work was the development of a diagnostic device for the use at the point-of-need for the detection of periodontal pathogens to enable a personalized therapy for treatment of periodontitis.

          Methods

          This test system is based on the polymerase chain reaction of DNA isolated from periodontal pathogens and was examined to precisely detect species-specific sequences on a rotating chip with lyophilized reagents for polymerase chain reaction. The preservation of the reagents was optimized to ensure their stability during the storage.

          Results

          In the current work, we have developed a model point-of-care device and showed a proof of concept. It requires low sample volume, is timesaving and can therefore facilitate early diagnosis and treatment of periodontal diseases.

          Conclusions

          The developed device can provide fast diagnosis of the composition and amount of patients’ oral flora and might help to assess the stage of periodontitis infection. This can facilitate an optimization of therapeutic approaches in order to prevent some of the more serious consequences of the disease.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12903-015-0155-y) contains supplementary material, which is available to authorized users.

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          Most cited references22

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          Point-of-care nucleic acid testing for infectious diseases.

          Nucleic acid testing for infectious diseases at the point of care is beginning to enter clinical practice in developed and developing countries; especially for applications requiring fast turnaround times, and in settings where a centralized laboratory approach faces limitations. Current systems for clinical diagnostic applications are mainly PCR-based, can only be used in hospitals, and are still relatively complex and expensive. Integrating sample preparation with nucleic acid amplification and detection in a cost-effective, robust, and user-friendly format remains challenging. This review describes recent technical advances that might be able to address these limitations, with a focus on isothermal nucleic acid amplification methods. It briefly discusses selected applications related to the diagnosis and management of tuberculosis, HIV, and perinatal and nosocomial infections. Copyright © 2011. Published by Elsevier Ltd.
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            rrndb: the Ribosomal RNA Operon Copy Number Database.

            The Ribosomal RNA Operon Copy Number Database (rrndb) is an Internet-accessible database containing annotated information on rRNA operon copy number among prokaryotes. Gene redundancy is uncommon in prokaryotic genomes, yet the rRNA genes can vary from one to as many as 15 copies. Despite the widespread use of 16S rRNA gene sequences for identification of prokaryotes, information on the number and sequence of individual rRNA genes in a genome is not readily accessible. In an attempt to understand the evolutionary implications of rRNA operon redundancy, we have created a phylogenetically arranged report on rRNA gene copy number for a diverse collection of prokaryotic microorganisms. Each entry (organism) in the rrndb contains detailed information linked directly to external websites including the Ribosomal Database Project, GenBank, PubMed and several culture collections. Data contained in the rrndb will be valuable to researchers investigating microbial ecology and evolution using 16S rRNA gene sequences. The rrndb web site is directly accessible on the WWW at http://rrndb.cme. msu.edu.
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              Comparison of the microbiota of supra- and subgingival plaque in health and periodontitis.

              The purpose of the present investigation was to compare the microbial composition of supra and subgingival plaque in 22 periodontally healthy (mean age 32+/-16 years) and 23 adult periodontitis subjects (mean age 51+/-14 years). A total of 2358 supra and separately subgingival plaque samples were collected from the mesial aspect of all teeth excluding 3rd molars in each subject. Samples were examined for the presence and levels of 40 bacterial taxa using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Clinical assessments including dichotomous measures of gingival redness, bleeding on probing, plaque accumulation and suppuration, as well as duplicate measures of pocket depth and attachment level, were made at 6 sites per tooth. Mean counts (x10(5), % DNA probe count and % sites colonized for each species were determined separately for supra and subgingival samples in each subject and then averaged across subjects in the 2 clinical groups. Significance of differences between healthy and periodontitis subjects was determined using the Mann-Whitney test and adjusted for multiple comparisons. Mean total DNA probe counts (x10(5), +/-SEM) for healthy and periodontitis subjects in supragingival plaque were 72.1+/-11 and 132+/-17.5, respectively (p<0.01), and in subgingival plaque 22.1+/-6.6 and 100.3+/-18.4, (p<0.001). Porphyromonas gingivalis, Bacteroides forsythus and Treponema denticola could be detected in supragingival plaque samples of both healthy and periodontitis subjects. Actinomyces species were the dominant taxa in both supra- and subgingival plaque from healthy and periodontitis subjects. 4 Actinomyces species accounted for 63.2%, of supragingival and 47.2% of subgingival plaque in healthy subjects and 48.% and 37.8% in periodontitis subjects respectively. Increased proportions of P. gingivalis, B. forsythus, and species of Prevotella, Fusobacterium, Campylobacter and Treponema were detected subgingivally in the periodontitis subjects. P. gingivalis, B. forsythus and T. denticola were significantly more prevalent in both supra- and subgingival plaque samples from periodontitis subjects. The main differences between supra and subgingival plaque as well as between health and disease were in the proportions and to some extent levels of Actinomyces, "orange" and "red" complex species.
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                Author and article information

                Contributors
                +49341355369326 , cornelia.gaertig@izi.fraunhofer.de
                Journal
                BMC Oral Health
                BMC Oral Health
                BMC Oral Health
                BioMed Central (London )
                1472-6831
                24 December 2015
                24 December 2015
                2015
                : 15
                : 165
                Affiliations
                [ ]Nanotechnology Unit, Department of Diagnostics, Fraunhofer Institute for Cell Therapy and Immunology, Perlickstraße 1, 04103 Leipzig, Germany
                [ ]Friedrich Schiller University, Jena, Germany
                [ ]Scanbec GmbH, Bitterfeld-Wolfen, Germany
                Article
                155
                10.1186/s12903-015-0155-y
                4690268
                29e22811-651d-4577-8fef-8bba026c2b2b
                © Gaertig et al. 2015

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 8 July 2015
                : 17 December 2015
                Funding
                Funded by: Federal Ministry for Economic Affairs and Energy
                Award ID: KF2302705AK1
                Categories
                Technical Advance
                Custom metadata
                © The Author(s) 2015

                Dentistry
                bacterial quantification,periodontitis,real-time pcr,microbial diagnostics,lyophilization

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