中文摘要：鉴定转基因水稻污染案例数量的增加可追溯到2006年的欧盟食品和饲料快速预警系统（RASFF）。由于可靠检测方法的不足，2011/884/EU决策提出用新的筛选方法来取代2008/289/EC决策，来找出所有原产于中国的转基因水稻。然而，2011/884/EU 提出的SYBR Green实时荧光定量PCR在目前的使用中缺少与其他TaqMan方法的一致性。为了评估2011/884/EU决策和2008/289/EC决策所提方法的特异性和可重复性，本文收集了原产于六个国家或地区的74种水稻，并用2011/884/EU决策和2008/289/EC决策所提方法对74种水稻样品进行了测试。研究发现，2011/884/EU决策提出的SYBR Green实时荧光定量PCR检测出24.3%（18 / 74）的样品呈阳性，而2011/884/EU决策和2008/289/EC决策提出的TaqMan探针实时PCR检测结果均为阴性。PCR扩增的ClylA基因序列表明，转基因水稻包括引物二聚体而非靶序列。由于出现高的假阳性结果，2011/884/EU决策提出的检测转基因水稻的SYBR Green实时荧光定量PCR需要改进。
外文摘要：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 ClylA(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 CrylA(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.
外文关键词：genetically modified organism; Decision 2011/884/EU; SYBR Green real-time PCR; false positive
作者：Xiao Qi-sheng; Xu Wen-tao; Yang Jie-lin; 等
作者单位：Shanghai Entry & Exit Inspect & Quarantine Bur
期刊名称：JOURNAL OF INTEGRATIVE AGRICULTURE
期刊影响因子：0.724
出版年份：2016
出版刊次：12
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