Journal of Integrative Agriculture ›› 2016, Vol. 15 ›› Issue (12): 2899-2910.DOI: 10.1016/S2095-3119(16)61378-3

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  • 收稿日期:2015-11-05 出版日期:2016-12-01 发布日期:2016-12-02

Evaluation of genetically modified rice detection methods 2011/884/ EU and 2008/289/EC proposed by the European Union

XIAO Qi-sheng1, 2, XU Wen-tao3, YANG Jie-lin1, PAN Liang-wen1   

  1. 1 Technical Center for Animal, Plant and Food Inspection and Quarantine, Shanghai Entry and Exit Inspection and Quarantine Bureau, Shanghai 200135, P.R.China
    2 College of Food Science & Technology, Shanghai Ocean University, Shanghai 201306, P.R.China
    3 Beijing Laboratory for Food Quality and Safety/College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R.China
  • Received:2015-11-05 Online:2016-12-01 Published:2016-12-02
  • Contact: YANG Jie-lin, Tel: +86-21-38620636, Fax: +86-21-68543124, E-mail: yangjl@shciq.gov.cn
  • About author:XIAO Qi-sheng, E-mail: xiaoqsheng@163.com
  • Supported by:

    This work was supported by the Science and Technology Project of Yangtze River Delta, China (16395810100), the National Transgenic Major Project, China (2012ZX08011003-1) and the Special Subject of Shanghai Technical Barriers to Trade, China (13TBT001).

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Abstract:     Increases in the number of cases of identified genetically modified (GM) rice contamination can be traced back to the first Rapid Alert System for Food and Feed (RASFF) in 2006. In response to the lack of reliable detection methods, Decision 2011/884/EU proposed that new screening methods replace Decision 2008/289/EC, to identify all possible GM rice products originating in China. However, the synergy brands (SYBR) Green real-time PCR assay proposed by Decision 2011/884/EU has been shown to lack conformity with other TaqMan methods currently in use. To evaluate the specificity and repeatability of the methods recommended in Decision 2011/884/EU and Decision 2008/289/EC, we collected 74 rice products originating from six countries or districts. The 74 rice samples were tested using the Decision 2011/884/EU and Decision 2008/289/EC methods. The parallel use of different instruments and reagents were used for testing in parallel, and the results were analyzed statistically. To avoid the limitations of specific laboratories, eight GM organism detection laboratories in China participated in a collaborative trial. In our tests, 24.3% (18/74) of the samples tested were positive with the SYBR Green real-time PCR assay using the Decision 2011/884/EU method, but were negative with the TaqMan real-time PCR assay using the Decision 2011/884/EU and Decision 2008/289/EC methods. Sequencing the PCR-amplified CryIA(b/c) genes in three samples (6, 30 and 43) showed that the products consisted of primer dimers rather than the targeted sequence. The combined experimental results showed that testing for the nopaline synthase gene (NOS) of Agrobacterium tumefasciens terminator and CryIA(b/c) produced false-positive results when the Decision 2011/884/EU method was used. Because of the high rate of false-positive results, the Decision 2011/884/EU SYBR Green method to detect GM rice requires improvement.

Key words: genetically modified organism ,  Decision 2011/884/EU ,  SYBR Green real-time PCR ,  false positive