Reference materials (RMs) for analysis of the human factor II (prothrombin) gene G20210A mutation.
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.
Klein
Christoph L
CL
Institute for Reference Materials and Measurements, European Commission, Joint Research Centre, Geel, Belgium. christoph.klein@cec.eu.int
Márki-Zay
János
J
Corbisier
Philippe
P
Gancberg
David
D
Cooper
Susan
S
Gemmati
Donato
D
Halbmayer
Walter-Michael
WM
Kitchen
Steve
S
Melegh
Béla
B
Neumaier
Michael
M
Oldenburg
Johannes
J
Leibundgut
Elisabeth Oppliger
EO
Reitsma
Pieter H
PH
Rieger
Sandra
S
Schimmel
Heinz G
HG
Spannagl
Michael
M
Tordai
Attilia
A
Tosetto
Alberto
A
Visvikis
Sophie
S
Zadro
Renata
R
Mannhalter
Christine
C
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
D017931:DNA Primers; D011516:Prothrombin; D004247:DNA