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Clinical Oral Investigations

, Volume 20, Issue 9, pp 2515–2528

First Online: 29 March 2016Received: 18 December 2014Accepted: 10 February 2016

Abstract

ObjectivesDespite the input of microbiome research, a group of 20 bacteria continues to be the focus of periodontal diagnostics and therapy. The aim of this study was to compare three commercial kits and laboratory-developed primer pairs for effectiveness in detecting such periodontopathogens.

Materials and methodsFourteen bacterial mock communities, consisting of 16 randomly assembled bacterial strains, were used as reference standard for testing kits and primers. Extracted DNA from mock communities was analyzed by PCR in-house with specific primers and forwarded for analysis to the manufacturer’s laboratory of each of the following kits: ParoCheck®Kit 20, micro-IDent®plus11, and Carpegen® Perio Diagnostik.

ResultsThe kits accurately detected Fusobacterium nucleatum, Prevotella intermedia-Prevotella nigrescens, Parvimonas micra, Aggregatibacter actinomycetemcomitans, Campylobacter rectus-showae, Streptococcus mitis, Streptococcus mutans, and Veillonella parvula. The in-house primers for F.nucleatum were highly specific to subtypes of the respective periopathogen. Other primers repeatedly detected oral pathogens not present in the mock communities, indicating reduced specificity.

ConclusionsThe commercial kits used in this study are reliable tools to support periodontal diagnostics. Whereas the detection profile of the kits is fixed at a general specificity level, the design of primers can be adjusted to differentiate between highly specific strains. In-house primers are more error-prone. Bacterial mock communities can be established as a reference standard for any similar testing.

Clinical relevanceThe tested kits render good results with selected bacterial species. Primers appear to be less useful for routine clinical diagnostics and of limited applicability in research. Basic information about the periodontopathogens identified in this study supports clinical decision-making.

KeywordsAggregatibacter actinomycetemcomitans Campylobacter rectus-showae Eikenella corrodens Fusobacterium nucleatum Microbiological diagnosis Mock community Molecular diagnostics Oral biofilm Parvimonas micra Periodontal diagnostics Prevotella intermedia Prevotella nigrescens Streptococcus mitis Streptococcus mutans Veillonella parvula  Download fulltext PDF



Autor: Elisabeth Santigli - Eva Leitner - Gernot Wimmer - Harald H. Kessler - Gebhard Feierl - Martin Grube - Katharina Eberhard

Fuente: https://link.springer.com/







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