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Licensed Unlicensed Requires Authentication Published by De Gruyter September 21, 2011

Reference materials (RMs) for analysis of the human factor II (prothrombin) gene G20210A mutation

  • Christoph L. Klein , János Márki-Zay , Philippe Corbisier , David Gancberg , Susan Cooper , Donato Gemmati , Walter-Michael Halbmayer , Steve Kitchen , Béla Melegh , Michael Neumaier , Johannes Oldenburg , Elisabeth Oppliger Leibundgut , Pieter H. Reitsma , Sandra Rieger , Heinz G. Schimmel , Michael Spannagl , Attilia Tordai , Alberto Tosetto , Sophie Visvikis , Renata Zadro and Christine Mannhalter

Abstract

The Scientific Committee of Molecular Biology Techniques (C-MBT) in Clinical Chemistry of the IFCC has initiated a joint project in co-operation with the European Commission, Joint Research Centre, Institute of Reference Materials and Measurements to develop and produce plasmid-type reference materials (RMs) for the analysis of the human prothrombin gene G20210A mutation. Although DNA tests have a high impact on clinical decision-making and the number of tests performed in diagnostic laboratories is high, issues of quality and quality assurance exist, and currently only a few RMs for clinical genetic testing are available. A gene fragment chosen was produced that spans all primer annealing sites published to date. Both the wild-type and mutant alleles of this gene fragment were cloned into a pUC18 plasmid and two plasmid RMs were produced. In addition, a mixture of both plasmids was produced to mimic the heterozygous genotype. The present study describes the performance of these reference materials in a commutability study, in which they were tested by nine different methods in 13 expert laboratories. This series of plasmid RMs are, to the best of our knowledge, the first plasmid-type clinical genetic RMs introduced worldwide.


Corresponding author: Christoph L. Klein, European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Retieseweg 111, 2440 Geel, Belgium Phone: +32-14-571634, Fax: +32-14-571548,

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Received: 2005-3-7
Accepted: 2005-6-17
Published Online: 2011-9-21
Published in Print: 2005-8-1

©2005 by Walter de Gruyter Berlin New York

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