title: Reference materials (RMs) for analysis of the human factor II (prothrombin) gene G20210A mutation.
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.
authors.affiliation: Institute for Reference Materials and Measurements, European Commission, Joint Research Centre, Geel, Belgium. christoph.klein@cec.eu.int
authors.forename: Christoph L
authors.initials: CL
authors.lastname: Klein
authors.forename: János
authors.initials: J
authors.lastname: Márki-Zay
authors.forename: Philippe
authors.initials: P
authors.lastname: Corbisier
authors.forename: David
authors.initials: D
authors.lastname: Gancberg
authors.forename: Susan
authors.initials: S
authors.lastname: Cooper
authors.forename: Donato
authors.initials: D
authors.lastname: Gemmati
authors.forename: Walter-Michael
authors.initials: WM
authors.lastname: Halbmayer
authors.forename: Steve
authors.initials: S
authors.lastname: Kitchen
authors.forename: Béla
authors.initials: B
authors.lastname: Melegh
authors.forename: Michael
authors.initials: M
authors.lastname: Neumaier
authors.forename: Johannes
authors.initials: J
authors.lastname: Oldenburg
authors.forename: Elisabeth Oppliger
authors.initials: EO
authors.lastname: Leibundgut
authors.forename: Pieter H
authors.initials: PH
authors.lastname: Reitsma
authors.forename: Sandra
authors.initials: S
authors.lastname: Rieger
authors.forename: Heinz G
authors.initials: HG
authors.lastname: Schimmel
authors.forename: Michael
authors.initials: M
authors.lastname: Spannagl
authors.forename: Attilia
authors.initials: A
authors.lastname: Tordai
authors.forename: Alberto
authors.initials: A
authors.lastname: Tosetto
authors.forename: Sophie
authors.initials: S
authors.lastname: Visvikis
authors.forename: Renata
authors.initials: R
authors.lastname: Zadro
authors.forename: Christine
authors.initials: C
authors.lastname: Mannhalter
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000032:analysis / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D017354:Point Mutation; D011516:Prothrombin / Q000235:genetics* / Q000592:standards*; D012015:Reference Standards
chemical_list: D017931:DNA Primers; D011516:Prothrombin; D004247:DNA