Guidance document on multiplex real-time PCR methodsGrohmann, Lutz (Avtor) Barbante, Alessandra (Avtor) Eriksson, Ronnie (Avtor) Gatto, Francesco (Avtor) Georgieva, Tzveta (Avtor) Huber, Ingrid (Avtor) Hulin, Julie (Avtor) Köppel, René (Avtor) Marchesi, Ugo (Avtor) Marmin, Lucas (Avtor) Mazzara, Marco (Avtor) Narendja, Frank (Avtor) Owen, Heather (Avtor) Perri, Elena (Avtor) Scholtens, Ingrid (Avtor) Sovová, Tereza (Avtor) Sowa, Slawomir (Avtor) Štebih, Dejan (Avtor) Weidner, Christopher (Avtor) Zdeňková, Kamila (Avtor) This document provides guidance for the development of new qualitative and quantitative multiplex real-time methods (mpPCR) and their implementation in a routine testing laboratory for food, feed and seeds. When implementing a mpPCR method, the laboratory should perform either an (in-house) validation or a verification of the method before its use. The various method performance parameters of the specific requirements for mpPCR are described and examples are given. Several different aspects have been considered, including the selection of the fluorophores, the master mix and the real-time PCR instrument used. The technical aspects are examined and recommendations are made on the choice of the detection and quantification strategy. Finally, a separate chapter is dedicated to troubleshooting (e.g. underperforming method, no or poor signal strength, cross-talk). In addition, a collection of existing mpPCR methods with emphasis on GMO detection is presented. Publications Office of the European Union20212024-09-02 12:06:44Temeljna raziskava20262UDK: 577ISBN: 978-92-76-37820-4DOI: 10.2760/243914COBISS_ID: 94024963sl