European Food Research and Technology

, Volume 227, Issue 6, pp 1621–1632

Validation of the performance of a GMO multiplex screening assay based on microarray detection

  • Serge Leimanis
  • Sandrine Hamels
  • Florence Nazé
  • Guillaume Mbongolo Mbella
  • Myriam Sneyers
  • Rupert Hochegger
  • Hermann Broll
  • Lillian Roth
  • Klára Dallmann
  • Adrienn Micsinai
  • José Luis La Paz
  • Maria Pla
  • Claudia Brünen-Nieweler
  • Nina Papazova
  • Isabel Taverniers
  • Norbert Hess
  • Britta Kirschneit
  • Yves Bertheau
  • Colette Audeon
  • Valérie Laval
  • Ulrich Busch
  • Sven Pecoraro
  • Katrin Neumann
  • Sibylle Rösel
  • Jeroen van Dijk
  • Esther Kok
  • Gianni Bellocchi
  • Nicoletta Foti
  • Marco Mazzara
  • William Moens
  • José Remacle
  • Guy Van Den Eede
Original Paper

DOI: 10.1007/s00217-008-0886-y

Cite this article as:
Leimanis, S., Hamels, S., Nazé, F. et al. Eur Food Res Technol (2008) 227: 1621. doi:10.1007/s00217-008-0886-y

Abstract

A new screening method for the detection and identification of GMO, based on the use of multiplex PCR followed by microarray, has been developed and is presented. The technology is based on the identification of quite ubiquitous GMO genetic target elements first amplified by PCR, followed by direct hybridisation of the amplicons on a predefined microarray (DualChip® GMO, Eppendorf, Germany). The validation was performed within the framework of a European project (Co-Extra, contract no 007158) and in collaboration with 12 laboratories specialised in GMO detection. The present study reports the strategy and the results of an ISO complying validation of the method carried out through an inter-laboratory study. Sets of blind samples were provided consisting of DNA reference materials covering all the elements detectable by specific probes present on the array. The GMO concentrations varied from 1% down to 0.045%. In addition, a mixture of two GMO events (0.1% RRS diluted in 100% TOPAS19/2) was incorporated in the study to test the robustness of the assay in extreme conditions. Data were processed according to ISO 5725 standard. The method was evaluated with predefined performance criteria with respect to the EC CRL method acceptance criteria. The overall method performance met the acceptance criteria; in particular, the results showed that the method is suitable for the detection of the different target elements at 0.1% concentration of GMO with a 95% accuracy rate. This collaborative trial showed that the method can be considered as fit for the purpose of screening with respect to its intra- and inter-laboratory accuracy. The results demonstrated the validity of combining multiplex PCR with array detection as provided by the DualChip® GMO (Eppendorf, Germany) for the screening of GMO. The results showed that the technology is robust, practical and suitable as a screening tool.

Keywords

GMO DualChip GMO Microarray Screening Multiplex Validation PCR Inter-laboratory ring trial 

Abbreviation

CRL

Community Reference Laboratory

EC

European Commission

PCR

Polymerase Chain Reaction

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Serge Leimanis
    • 1
  • Sandrine Hamels
    • 1
  • Florence Nazé
    • 2
  • Guillaume Mbongolo Mbella
    • 2
  • Myriam Sneyers
    • 2
  • Rupert Hochegger
    • 3
  • Hermann Broll
    • 4
  • Lillian Roth
    • 4
  • Klára Dallmann
    • 5
  • Adrienn Micsinai
    • 5
  • José Luis La Paz
    • 6
  • Maria Pla
    • 6
  • Claudia Brünen-Nieweler
    • 7
  • Nina Papazova
    • 8
  • Isabel Taverniers
    • 8
  • Norbert Hess
    • 9
  • Britta Kirschneit
    • 9
  • Yves Bertheau
    • 10
  • Colette Audeon
    • 10
  • Valérie Laval
    • 10
  • Ulrich Busch
    • 12
  • Sven Pecoraro
    • 12
  • Katrin Neumann
    • 13
  • Sibylle Rösel
    • 13
  • Jeroen van Dijk
    • 14
  • Esther Kok
    • 14
  • Gianni Bellocchi
    • 11
  • Nicoletta Foti
    • 11
  • Marco Mazzara
    • 11
  • William Moens
    • 11
  • José Remacle
    • 1
  • Guy Van Den Eede
    • 11
  1. 1.Eppendorf Array TechnologiesNamurBelgium
  2. 2.ISP, Institut Scientifique de Santé PubliqueBrusselsBelgium
  3. 3.AGES, Österreichische Agentur für Gesundheit und ErnährungssicherheitViennaAustria
  4. 4.BfR, Bundesinstitut für RisikobewertungBerlinGermany
  5. 5.BIOMI Ltd, BIOMI Ltd.GödöllõHungary
  6. 6.CSIC, Consejo Superior de Investigaciones CientíficasBarcelonaSpain
  7. 7.CVUA NRW, Chemisches Landes- und Staatliches VeterinäruntersuchungsamtMünsterGermany
  8. 8.DVP-ILVO, Instituut voor Landbouw- en VisserijonderzoekMerelebekeBelgium
  9. 9.IHU, Institut für Hygiene und UmweltHamburgGermany
  10. 10.INRA, Institut National de la Recherche AgronomiqueVersaillesFrance
  11. 11.European Commission, Joint Research Centre, Institute for Health and Consumer ProtectionBiotechnology and GMOs UnitIspraItaly
  12. 12.LGL, Bayer. Landesamt für Gesundheit und LebensmittelsicherheitOberschleißheimGermany
  13. 13.LIFEPRINT, Lifeprint GmbHIllertissenGermany
  14. 14.RIKILT, Institute of Food SafetyWageningen University and Research CentreWageningenThe Netherlands

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