title: Development and validation of duplex, triplex, and pentaplex real-time PCR screening assays for the detection of genetically modified organisms in food and feed.
abstract: Worldwide, qualitative methods based on PCR are most commonly used as screening tools for genetically modified material in food and feed. However, the increasing number and diversity of genetically modified organisms (GMO) require effective methods for simultaneously detecting several genetic elements marking the presence of transgenic events. Herein we describe the development and validation of a pentaplex, as well as complementary triplex and duplex real-time PCR assays, for the detection of the most common screening elements found in commercialized GMOs: P-35S, T-nos, ctp2-cp4-epsps, bar, and pat. The use of these screening assays allows the coverage of many GMO events globally approved for commercialization. Each multiplex real-time PCR assay shows high specificity and sensitivity with an absolute limit of detection below 20 copies for the targeted sequences. We demonstrate by intra- and interlaboratory tests that the assays are robust as well as cost- and time-effective for GMO screening if applied in routine GMO analysis.
authors: Bavarian Health and Food Safety Authority (LGL) , Veterinärstrasse 2, D-85764 Oberschleissheim, Germany.
authors: Ingrid
authors: I
authors: Huber
authors: Annette
authors: A
authors: Block
authors: Daniela
authors: D
authors: Sebah
authors: Frédéric
authors: F
authors: Debode
authors: Dany
authors: D
authors: Morisset
authors: Lutz
authors: L
authors: Grohmann
authors: Gilbert
authors: G
authors: Berben
authors: Dejan
authors: D
authors: Stebih
authors: Mojca
authors: M
authors: Milavec
authors: Jana
authors: J
authors: Zel
authors: Ulrich
authors: U
authors: Busch
mesh_terms: D000821:Animal Feed / Q000032:analysis*; D017931:DNA Primers / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D005506:Food Contamination / Q000032:analysis; D030841:Food, Genetically Modified; D030821:Plants, Genetically Modified / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D018744:DNA, Plant