title: The characterization and certification of a quantitative reference material for Legionella detection and quantification by qPCR.
abstract: OBJECTIVE
  The characterization and certification of a Legionella DNA quantitative reference material as a primary measurement standard for Legionella qPCR.
  
  
  RESULTS
  Twelve laboratories participated in a collaborative certification campaign. A candidate reference DNA material was analysed through PCR-based limiting dilution assays (LDAs). The validated data were used to statistically assign both a reference value and an associated uncertainty to the reference material.
  
  
  CONCLUSIONS
  This LDA method allowed for the direct quantification of the amount of Legionella DNA per tube in genomic units (GU) and the determination of the associated uncertainties. This method could be used for the certification of all types of microbiological standards for qPCR.
  
  
  CONCLUSIONS
  The use of this primary standard will improve the accuracy of Legionella qPCR measurements and the overall consistency of these measurements among different laboratories. The extensive use of this certified reference material (CRM) has been integrated in the French standard NF T90-471 (April 2010) and in the ISO Technical Specification 12 869 (Anon 2012 International Standardisation Organisation) for validating qPCR methods and ensuring the reliability of these methods.
authors: Hospices Civils de Lyon, Centre National de Référence des Légionelles, Lyon, France.
authors: M
authors: M
authors: Baume
authors: L
authors: L
authors: Garrelly
authors: J P
authors: JP
authors: Facon
authors: S
authors: S
authors: Bouton
authors: P O
authors: PO
authors: Fraisse
authors: C
authors: C
authors: Yardin
authors: M
authors: M
authors: Reyrolle
authors: S
authors: S
authors: Jarraud
mesh_terms: D002568:Certification; D007875:Legionella / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D012015:Reference Standards