The characterization and certification of a quantitative reference material for Legionella detection and quantification by qPCR
| Autor: | M. Baume, C. Yardin, L. Garrelly, M. Reyrolle, S. Bouton, S. Jarraud, J.P. Facon, P.O. Fraisse |
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| Přispěvatelé: | Laboratoire de Physique Corpusculaire - Clermont-Ferrand (LPC), Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Certification
Legionella [SDV]Life Sciences [q-bio] Real-Time Polymerase Chain Reaction Bioinformatics Applied Microbiology and Biotechnology MESH: Certification 03 medical and health sciences Limiting dilution Reliability (statistics) 030304 developmental biology 0303 health sciences biology 030306 microbiology MESH: Real-Time Polymerase Chain Reaction General Medicine Reference Standards biology.organism_classification Reliability engineering Certified reference materials Primary standard MESH: Reference Standards Biotechnology MESH: Legionella |
| Zdroj: | Journal of Applied Microbiology Journal of Applied Microbiology, Wiley, 2013, 114 (6), pp.1725-33. ⟨10.1111/jam.12172⟩ Journal of Applied Microbiology, 2013, 114 (6), pp.1725-33. ⟨10.1111/jam.12172⟩ |
| ISSN: | 1364-5072 1365-2672 |
| DOI: | 10.1111/jam.12172⟩ |
| Popis: | Aims The characterization and certification of a Legionella DNA quantitative reference material as a primary measurement standard for Legionella qPCR. Methods and 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. Significance and Impact of the Study 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. |
| Databáze: | OpenAIRE |
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