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|Title:||Validation of the Performance of a GMO Multiplex Screening Assay based on Microarray Detection|
|Authors:||LEIMANIS Serge; HAMELS Sandrine; NAZÉ Florence; SNEYERS Myriam; MBONGOLO MBELLA Guillaume; HOCHEGGER Rupert; BROLL Hermann; ROTH Lillian; DALLMANN Klara; MICSINAI Adrienn; LA PAZ José Luis; PLA Maria; BRUNEN-NIEWELER Claudia; PAPAZOVA Nina; TAVERNIERS Isabelle; HESS Norbert; KIRSCHNEIT Britta; BERTHEAU Yves; AUDÉON Colette; LAVAL Valerie; BUSCH Ulrich; PECORARO Sven; NEUMANN Katrin; ROSEL Sibylle; VAN DIJK Jeroen P.; KOK Esther J.; BELLOCCHI GIANNI; FOTI NICOLETTA; MAZZARA MARCO; MOENS WILLIAM; REMACLE José; VAN DEN EEDE GUY|
|Citation:||EUROPEAN FOOD RESEARCH AND TECHNOLOGY vol. 227 no. 6 p. 1621-1632|
|JRC Publication N°:||JRC42558|
|Type:||Articles in Journals|
|Abstract:||A new multiple targets screening method for the detection and identification of GMO based on the use of multiplex PCR followed by microarray, has been developed for the detection and identification of GMO and is presented in the accompanying paper. 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 twelve laboratories specialised in GMO detection. The present article reports the strategy and the results of an ISO 5725 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 at concentration of 0.1 and 99.9 % was incorporated in the study to test for 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 demonstrate 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.|
|JRC Institute:||Institute for Health and Consumer Protection|
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