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Record ID
Date of creation
2021-03-23 15:07 UTC (austein.mcloughlin@cbd.int)
Date of publication
2021-03-23 15:07 UTC (austein.mcloughlin@cbd.int)

General Information
Development and comparative study of a pat/ bar real-time PCR assay for integrating the screening strategy of a GMO testing laboratory
Daniela Verginelli, Annalisa Paternò, Maria Laura De Marchis, Cinzia Quarchioni, Daniela Vinciguerra, Pamela Bonini, Stefania Peddis, Cristiana Fusco, Marisa Misto, Cristina Marfoglia, Francesco Pomilio, Ugo Marchesi
Author’s contact information
Ugo Marchesi, Unità Operativa Semplice a valenza Direzionale - Ricerca e controllo degli organismi geneticamente modificati Centro di referenza nazionale per la ricerca di organismi geneticamente  modificati (CROGM) Laboratorio Nazionale di Riferimento per alimenti e mangimi geneticamente modificati National Reference Laboratory for GM food and feed Via Appia Nuova 141100178 Roma - Italia

Kindly refer to the attached article for contact's email address
  • English
Publication date
Summary, abstract or table of contents
BACKGROUND: The number and variety of genetically modified organisms (GMOs) used globally for the production of food and feed, and potentially circulating in the European Union (EU), is constantly increasing. This implies an additional effort for the EU enforcement laboratories to optimize available resources, to contain costs and time. A well established approach for streamlining the analytical workflow is the introduction of a screening step, typically based on a smart set of real-time polymerase chain reaction (PCR) screening  methods. The multiplexing strategy, allowing the detection of several screening elements simultaneously, is a further optimization of this step.

RESULTS: In this study, we present the validation of a real-time PCR duplex assay for the pat and bar screening elements to be easily incorporated in the GMO diagnostic routine. We also provide a comparison between this method and the related singleplex and pre-spotted assays.

CONCLUSION: Our results fully respect all the validation parameters suggested by the Minimum Performance Criteria of the European Network of GMO Laboratories. Furthermore, the duplex assay is equivalent in terms of performance compared to the other two methods, but it shows a higher overall flexibility and cost effectiveness.
Thematic areas
Information on Organisms or LMOs
Gene(s) identification
Phosphinothricin N-acetyltransferase gene - Streptomyces hygroscopicus - STRHY
Resistance to herbicides - Glufosinate
Phosphinothricin N-acetyltransferase gene - Streptomyces viridochromogenes - STRVR
Resistance to herbicides - Glufosinate
Additional Information
Type of resource
  • Article (journal / magazine / newspaper)
DOI 10.1002/jsfa.10235
Publisher and its location
Journal of the Science of Food and Agriculture
John Wiley & Sons Ltd on behalf of Society of Chemical Industry
Open access
Journal article
Keywords and any other relevant information
LMO; multiplex real-time PCR; screening; pre-spotted plates; pat gene; bar gene; droplet digital PCR; ddPCR; transferability