512 Chapter 29
FAPAS ® - FEPAS ® - GeMMa (Food Analysis
Performance Assessment Scheme, Central
Science Laboratory, York, UK) promote
rounds of quantitative profi ciency testing and
qualitative (presence/absence) detection, in
which any food laboratory is encouraged to
participate to test its performance.
A common set of criteria for performance
of GMO detection methods should be evalu-
ated (Bertheau et al. 2002 ; Bellocchi et al.
2008 ; Ž el et al. 2008 ). Recommendations for
the validation of quantitative PCR methods
are presented by H ü bner and coworkers
(2001) , and currently there is an international
standard that defi nes the PCR performance
parameters for GMO analysis (ISO 2006 ).Implementation of Methodologies in
the Food and Feed Chains
GMO quality - control programs are increas-
ingly applied throughout food - chain produc-
tion under the framework of legislative
control measures. Thus, the availability of
reliable, rapid, and accepted test systems to
detect the presence or absence, or even the
degree of contamination of GMO, becomes
increasingly important for the agricultural
and food industry. However, the implementa-
tion of GMO detection methodologies in the
food and animal feed chains is not a simple
issue. For a realistic implementation in food
laboratories, several important aspects, such
as the sampling procedure and the use of
adequate reference materials for controls and
standards, as well as the selection of the ana-
lytical methods, must be seriously consid-
ered. In addition, in the GMO analysis
scenario, two main limitations are still
present: (1) the important restrictions for
access to protected transgene sequences, and
(2) the scarce availability of GM certifi ed
material usually provided by the biotech
companies only under strict confi dentiality
agreements. In this context, nowadays, certi-
fi ed reference material (CRM) is only avail-
able for the following GMOs: GTS40 - 3 - 2,cost - effective, reliable, and having a wider
linear range of detection compared with
genomic reference material
A new strategy for construction of refer-
ence material has been devised (Roth et al.
2008 ). This technique is based on the genome
amplifi cation by multiple displacement
amplifi cation (MDA). It is based on the use
of a specifi c DNA polymerase, phi29 DNA
polymerase, and random hexamer primers
for the replication of genomic DNA in an
isothermal reaction at 30 ° C, leading to the
synthesis of large amounts of DNA with
fragments > 70 kb in size.
Validation of Analytical Methods
Methods for the detection, identifi cation, and
quantifi cation of GMOs have become widely
used by enforcement laboratories, and the
number of published systems has increased
considerably in recent years. Nowadays, a
wide range of methods is available, which
can generate confusion for fi nal users. In
order to determine the suitability of each
method for providing reliable analytical data,
every method should be validated (Anklam
et al. 2002 ). The concept of validation implies
that the application of a given method must
provide similar analytical results in different
laboratories using different reagents and
operators. For this reason, the validation
process must involve several laboratories,
which have to be coordinated by a principal
one, usually different from the laboratory
that developed the method. In Europe, there
are institutes that coordinate GMO validation
studies, such as the Federal Institute for
Health Protection of Consumers and
Veterinary Medicine (BgVV) in Germany,
or the Community Reference Laboratory
(CRL) through the Biotechnology and GMOs
Unit of the Joint Research Centre (JRC)
in Italy ( http://gmo-crl.jrc.it ). The Gipsa
Grain Inspection, Packers and Stockyards
Administration (GIPSA) of the U.S.
Department of Agriculture (USDA) and