Guidance document on multiplex real-time PCR methods
European Network of GMO Laboratories
Polymerase Chain Reaction (PCR) is considered the gold standard technique for the detection and quantification of Genetically Modified (GM) food, feed and seeds. The combination of two or more PCR tests in a single reaction tube is referred to as multiplex PCR and is a powerful approach already adopted by many testing laboratories. Multiplex PCR can address the need for increased and more efficient testing for the presence of GMOs in food, feed or seeds and for the quantification of the GM event.
Although real-time PCR has been used for GMO testing for many years as well as for research, forensic or diagnostic purposes, a comprehensive guidance on the development and implementation of multiplex PCR is currently not available.
This document has been developed by experts from the European Network of GMO Laboratories (ENGL). It provides background information on the technical aspects of multiplex PCR (mpPCR), including information on instruments, fluorescent dye selection and reaction controls. It gives guidance on how to implement a previously validated mpPCR method (i.e. method verification), as well as how to develop and validate a new mpPCR method.
Guidance on criteria for the acceptance of methods, examples of how to assess these criteria and troubleshooting tips are given. Finally, a list of currently used mpPCR methods is included.
GROHMANN Lutz;
BARBANTE A.;
ERIKSSON Ronnie;
GATTO Francesco;
GEORGIEVA Tzveta;
HUBER Ingrid;
HULIN Julie;
KÖPPEL Renè;
MARCHESI Ugo;
MARMIN Lucas;
MAZZARA Marco;
NARENDJA Frank;
OWEN Heather;
PERRI Elena;
SCHOLTENS Ingrid;
SOVOVÁ Tereza;
SOWA Slawomir;
STEBIH Dejan;
WEIDNER Christopher;
ZDEŇKOVÁ Kamila;
2021-08-13
Publications Office of the European Union
JRC125188
978-92-76-37820-4 (online),
1831-9424 (online),
EUR 30708 EN,
OP KJ-NA-30708-EN-N (online),
https://publications.jrc.ec.europa.eu/repository/handle/JRC125188,
10.2760/243914 (online),
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