Event-specific Method for the Quantification of Maize MIR162 by Real-time PCR

The European Union Reference Laboratory for Genetically Modified Food and Feed (EURL-GMFF), established by Regulation (EC) No 1829/2003, in collaboration with the European Network of GMO Laboratories (ENGL), has carried out a collaborative study to assess the performance of a quantitative event-specific method to detect and quantify the MIR162 transformation event (unique identifier SYN-IR162-4) in maize DNA. The collaborative study was conducted according to internationally accepted guidelines (1, 2). In accordance to Regulation (EC) No 1829/2003 of 22 September 2003 on genetically modified food and feed and with Regulation (EC) No 641/2004 of 6 April 2004 on detailed rules for the implementation of Regulation (EC) No 1829/2003, Syngenta Seeds S.A.S. provided the detection method and the control samples (genomic DNA extracted from homogenised seeds containing the transformation event and from conventional homogenised seeds). The EURL-GMFF prepared the validation samples (calibration samples and blind samples at unknown GM percentage [DNA/DNA]). The collaborative study involved twelve laboratories from nine European countries. The results of the international collaborative study met the ENGL performance requirements. The method is therefore considered applicable to the control samples provided, in accordance with the requirements of Annex I-2.C.2 to Commission Regulation (EC) No 641/2004. The results of the collaborative study are made publicly available at http://gmo-crl.jrc.ec.europa.eu/.JRC.I.3-Molecular Biology and Genomic

Report on the Verification of the Performance of Bt11, MIR162, MIR604 and GA21 Event-specific PCR-based Methods Applied to DNA Extracted from Stack Maize Bt11 x MIR162 x MIR604 x GA21

An application was submitted by Syngenta Crop Protection AG to request the authorisation of genetically modified Bt11 x MIR162 x MIR604 x GA21 maize (resistant to lepidopteran and coleopteran pests, able to utilise mannose as the only primary carbon source and tolerant to glufosinate ammonium and glyphosate) and all sub-combinations of the individual events as present in the segregating progeny, for food and feed uses, and import and processing, in accordance with articles 5 and 17 of Regulation (EC) N° 1829/2003 GM Food and GM Feed. The unique identifier assigned to Bt11 x MIR162 x MIR604 x GA21 maize is SYN-BTØ11-1x YN-IR162-4xSYN-IR6Ø4-5xMON-ØØØ21-9. The genetically modified maize line Bt11 x MIR162 x MIR604 x GA21 maize has been obtained by conventional crossing of four genetically modified maize events: Bt11, MIR162, MIR604, and GA21 without any new genetic modification. The EU-RL GMFF has previously validated individually, and declared fit for purpose, the detection methods for the single events Bt11, MIR162, MIR604 and GA21 (see http://gmo-crl.jrc.ec.europa.eu/StatusOfDossiers.aspx). In line with the approach defined by the ENGL (http://gmo-crl.jrc.ec.europa.eu/doc/Min_Perf_Requirements_Analytical_methods.pdf) the EU-RL GMFF therefore has carried out only an in-house verification of the performance of each validated method when applied to DNA extracted from Bt11 x MIR162 x MIR604 x GA21. The hereby reported the in-house verification study lead to the conclusion that the individual methods meet the ENGL criteria also when applied to DNA extracted from the GM maize stack Bt11 x MIR162 x MIR604 x GA21JRC.I.3-Molecular Biology and Genomic

Comparative Testing Report on the Detection and Quantification of Maize Event MON 810 - Comparative testing round: ILC-CRL-GMFF-CT-02/10

In the frame of Regulation (EC) No 882/2004, the European Union Reference Laboratory for Genetically Modified Food and Feed has the duty to organise comparative testing rounds and to ensure an appropriate follow-up of these activities. This report describes the outcome of the second comparative testing round ILC-CRL-GMFF-CT-02/10. Participants had to determine the GM content in two test items denoted maize powder levels 1 and 2, containing different GM percentages of maize event MON 810. This comparative testing round was organised in collaboration with the Reference Materials Unit and the Food Safety and Quality Unit of the Institute for Reference Materials and Measurements (Geel, BE). The maize event MON 810 test items were produced by the Reference Materials Unit. The Food Safety and Quality Unit managed the on-line registration and submission of results. A total of 136 laboratories were invited to participate in ILC-CRL-GMFF-CT-02/10. Six National Reference Laboratories declined participation, of which two were no longer a National Reference Laboratory. Ninety laboratories from 41 countries returned results, of which 65 were National Reference Laboratories, six were members of the European Network of GMO Laboratories only and 19 were laboratories from third countries. Two National Reference Laboratories, two Official control laboratories and nine laboratories from a third country did not submit any results. Participants could report the results of the exercise either in mass/mass % or in copy/copy %. The outcome of this second comparative testing round was in general positive, with 82-100 % of participants gaining a z-score in the range of -2 to +2 for both maize powder levels 1 and 2 regardless of the calibration method, the measurement unit and the approach used for calculating the z-score.JRC.I.3-Molecular Biology and Genomic

Event-specific Method for the Quantification of Soybean Line MON 89788 Using Real-time PCR v. 1.01: Validation Report and Validated Method

The JRC as European Union Reference Laboratory for GM Food and Feed (EURL-GMFF), established by Regulation (EC) No 1829/2003, in collaboration with the European Network of GMO Laboratories (ENGL), has carried out a collaborative study to assess the performance of a quantitative event-specific method to detect and quantify the MON89788 transformation event in soybean DNA (unique identifier MON-89788-1). The collaborative trial was conducted according to internationally accepted guidelines (1, 2). In accordance with Regulation (EC) No 1829/2003 of 22 September 2003 on genetically modified food and feed and with Regulation (EC) No 641/2004 of 6 April 2004 on detailed rules for the implementation of Regulation (EC) No 1829/2003, Monsanto provided the detection method and the samples (soybean seeds containing the transformation event and conventional soybean seeds). The JRC prepared the validation samples (calibration samples and blind samples at unknown GM percentage [DNA/DNA]). The collaborative trial involved twelve laboratories from eight European countries. The results of the international collaborative trial met the ENGL performance requirements. The method is therefore considered applicable to the control samples provided, in accordance with the requirements of Annex I-2.C.2 to Commission Regulation (EC) No 641/2004.JRC.I.3-Molecular Biology and Genomic

HLA-DQB1 genotyping in a family with narcolepsy-cataplexy

Narcolepsy is a unique model for dysfunction in mechanisms that regulate sleep and wakefulness. the narcolepsy syndrome is characterized by excessive daytime sleepiness with recurrent episodes of irresistible sleep, cataplexy, hypnagogic and/or hypnopompic hallucinations and sleep paralysis. the current hypothesis for the etiology of narcolepsy is that it is an autoimmune disorder because of its strong association with the human leukocyte antigen (HLA) system. HLA-DQ alleles are not particularly mutated in narcoleptic patients but they directly influence susceptibility to the disease. DQB1(*)0602 homozygote carriers have a two to four times higher risk of developing the disease than heterozygote carriers. in the present study we report a rare multiplex familial case of narcolepsy-cataplexy and show the strong effect of the HLA-DQB1(*)0602 allele upon the disease phenotype. in the family studied herein, both the proband and his brother are severely affected and homozygous DQB1*0602, whereas their sister does not carry the allele and is not affected at all. These data corroborate previous findings proposing DQB1*0602 homozygous subjects to be far more susceptible to narcolepsy. Insights into the DQB1*0602 positive family that include homozygous subjects may prove to be an important asset in the investigation of genetic vs. environmental factors predisposing to narcolepsy. (c) 2007 Elsevier B.V. All rights reserved.Universidade Federal de São Paulo, Dept Psychobiol, São Paulo, BrazilUniv Chile, INTA, Santiago, ChileUniversidade Federal de São Paulo, Dept Psychobiol, São Paulo, BrazilWeb of Scienc