title: Testing the interaction between analytical modules: an example with Roundup Ready soybean line GTS 40-3-2.
abstract: BACKGROUND
  The modular approach to analysis of genetically modified organisms (GMOs) relies on the independence of the modules combined (i.e. DNA extraction and GM quantification). The validity of this assumption has to be proved on the basis of specific performance criteria.
  
  
  RESULTS
  An experiment was conducted using, as a reference, the validated quantitative real-time polymerase chain reaction (PCR) module for detection of glyphosate-tolerant Roundup Ready(R) GM soybean (RRS). Different DNA extraction modules (CTAB, Wizard and Dellaporta), were used to extract DNA from different food/feed matrices (feed, biscuit and certified reference material [CRM 1%]) containing the target of the real-time PCR module used for validation. Purity and structural integrity (absence of inhibition) were used as basic criteria that a DNA extraction module must satisfy in order to provide suitable template DNA for quantitative real-time (RT) PCR-based GMO analysis. When performance criteria were applied (removal of non-compliant DNA extracts), the independence of GMO quantification from the extraction method and matrix was statistically proved, except in the case of Wizard applied to biscuit. A fuzzy logic-based procedure also confirmed the relatively poor performance of the Wizard/biscuit combination.
  
  
  CONCLUSIONS
  For RRS, this study recognises that modularity can be generally accepted, with the limitation of avoiding combining highly processed material (i.e. biscuit) with a magnetic-beads system (i.e. Wizard).
authors.affiliation: European Commission, Joint Research Centre, Institute for Health and Consumer Protection, Molecular Biology and Genomics Unit, via E, Fermi 2749, 21027 Ispra, VA, Italy.
authors.forename: Gianni
authors.initials: G
authors.lastname: Bellocchi
authors.forename: Marzia
authors.initials: M
authors.lastname: De Giacomo
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Foti
authors.forename: Marco
authors.initials: M
authors.lastname: Mazzara
authors.forename: Eleonora
authors.initials: E
authors.lastname: Palmaccio
authors.forename: Cristian
authors.initials: C
authors.lastname: Savini
authors.forename: Chiara
authors.initials: C
authors.lastname: Di Domenicantonio
authors.forename: Roberta
authors.initials: R
authors.lastname: Onori
authors.forename: Guy
authors.initials: G
authors.lastname: Van den Eede
mesh_terms: D000821:Animal Feed / Q000032:analysis; D018744:DNA, Plant / Q000032:analysis / Q000302:isolation & purification; D005504:Food Analysis / Q000379:methods*; D030841:Food, Genetically Modified; D010865:Pilot Projects; D030821:Plants, Genetically Modified / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013025:Glycine max / Q000235:genetics*
chemical_list: D018744:DNA, Plant