Guidance document on multiplex real-time PCR methods (Aug 2021) | BCH-VLR-SCBD-116269 | Biosafety Virtual Library Resources | Biosafety Clearing-House


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last updated: 30 Aug 2021

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Guidance document on multiplex real-time PCR methods
Lutz Grohmann, A. Barbante, Ronnie Eriksson, Francesco Gatto, Tzveta Georgieva, Ingrid Huber, Julie Hulin, Renè Köppel, Ugo Marchesi, Lucas Marmin, Marco Mazzara, Frank Narendja, Heather Owen, Elena Perri, Ingrid Scholtens, Tereza Sovová, Slawomir Sowa, Dejan Stebih, Christopher Weidner, Kamila Zdeňková
European Commission - Joint Research Centre, Institute for Health and Consumer Protection, Unit Molecular Biology and Genomics (JRC)   BCH-ORG-SCBD-16002-14
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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.
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ISBN: 978-92-76-37820-4; ISSN: 1831-9424; EUR 30708 EN; OP KJ-NA-30708-EN-N; doi:10.2760/243914
Keywords and any other relevant information
technical reports; agriculture; safety; standards; PCR; multiplex pCR; mpPCR
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