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.