title: Accuracy of commercial kits and published primer pairs for the detection of periodontopathogens.
abstract: OBJECTIVE
  Despite 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.
  
  
  METHODS
  Fourteen 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.
  
  
  RESULTS
  The 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.
  
  
  CONCLUSIONS
  The 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.
  
  
  CONCLUSIONS
  The 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.
authors.affiliation: Division of Orthodontics and Maxillofacial Orthopedics, Department of Dentistry and Maxillofacial Surgery, Medical University of Graz, Billrothgasse 4, A-8010, Graz, Austria. elisabeth.santigli@medunigraz.at.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Santigli
authors.affiliation: Bacteriology and Mycology Laboratory, Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Universitätsplatz 4, 8010, Graz, Austria.
authors.forename: Eva
authors.initials: E
authors.lastname: Leitner
authors.affiliation: Division of Prosthodontics, Restaurative Dentistry, Periodontology and Implantology, Department of Dentistry and Maxillofacial Orthopedics, Medical University of Graz, Billrothgasse 4, 8010, Graz, Austria.
authors.forename: Gernot
authors.initials: G
authors.lastname: Wimmer
authors.affiliation: Research Unit Molecular Diagnostics, Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Universitätsplatz 4, 8010, Graz, Austria.
authors.forename: Harald H
authors.initials: HH
authors.lastname: Kessler
authors.affiliation: Bacteriology and Mycology Laboratory, Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Universitätsplatz 4, 8010, Graz, Austria.
authors.forename: Gebhard
authors.initials: G
authors.lastname: Feierl
authors.affiliation: Institute of Plant Sciences, University of Graz, Holteigasse 6, 8010, Graz, Austria.
authors.forename: Martin
authors.initials: M
authors.lastname: Grube
authors.affiliation: Center of Medical Research, Medical University of Graz, Stiftingtalstraße 24/1, 8010, Graz, Austria.
authors.forename: Katharina
authors.initials: K
authors.lastname: Eberhard
authors.affiliation: Division of Orthodontics and Maxillofacial Orthopedics, Department of Dentistry and Maxillofacial Surgery, Medical University of Graz, Billrothgasse 4, A-8010, Graz, Austria.|Institute of Plant Sciences, University of Graz, Holteigasse 6, 8010, Graz, Austria.
authors.forename: Barbara
authors.initials: B
authors.lastname: Klug
mesh_terms: D001431:Bacteriological Techniques; D004269:DNA, Bacterial / Q000032:analysis*; D006801:Humans; D010510:Periodontal Diseases / Q000382:microbiology*; D016133:Polymerase Chain Reaction
chemical_list: D004269:DNA, Bacterial
keywords: Aggregatibacter 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