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首页> 外文期刊>Quality Assurance and Safety of Crops & Foods >Single laboratory method performance evaluation for the analysis of Roundup Ready (R) soy flour by qualitative and quantitative detection methods
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Single laboratory method performance evaluation for the analysis of Roundup Ready (R) soy flour by qualitative and quantitative detection methods

机译：单一实验室绩效评价方法分析抗农达大豆(R)的面粉通过定性和定量检测方法

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摘要

The Republic of Turkey has approved 7 soybean and 25 corn genetically modified (GM) events for animal feed use only and the biosafety legislation has banned the cultivation of GM crops and requires that all genetically modified organisms (GMOs), including imports, to be approved for use and further establishes a strict policy of testing for food, feed and seed potentially containing GMOs. For the GMO analysis, each laboratory should establish the verification on method performance criteria and calculation of measurement uncertainty. The aim of this study is to define the verification of qualitative and quantitative detection of Roundup Ready (R) soybean as a model for single laboratory verification in the context of the European Network of GMOs Laboratories guidance documents. First, two methods were used for the extraction of nucleic acids (DNA) and their extraction efficiencies were compared based on the quantity, purity, fragmentation state of DNA and inhibition in polymerase chain reaction (PCR). Second, a verification procedure of a real time PCR method for qualitative detection of cauliflower mosaic virus 35S promoter and Agrobacterium tumefaciens Tnos sequences in DNA samples extracted from certified reference materials and GM soy flour samples was performed. Last, the standard curves were prepared in order to explain verification of quantitative real-time PCR analysis by reaching the ideal value of -3.62 for lec reference gene and -3.40 for A. tumefaciens strain CP4 5-enolpyruvulshikimate-3-phosphate synthase (EPSPS) enzyme target gene. All method performance criteria for quantification (within-laboratory reproducibility standard deviation, relative standard deviation, uncertainty, bias, limit of detection, limit of quantification, and linearity) were met and thus the method in this study was verified. Finally, the document highlights a clear example for analysis of GMOs in food and feed samples, and points out the need for interlaboratory studies at the national and international level.
机译：土耳其共和国7大豆和批准25玉米转基因(GM)事件动物饲料使用生物安全立法禁止种植转基因并要求所有转基因作物生物(GMOs),包括进口批准使用,进一步建立了一个严格的政策的测试食品、饲料和种子可能含有转基因生物。分析,每个实验室都应该建立验证性能标准和方法计算测量的不确定性。本研究定义的验证综述的定性和定量检测准备好(R)大豆作为单一的模型实验室验证的上下文欧洲的转基因生物实验室网络指导文档。提取的核酸(DNA)和他们的提取效率比较的基础上数量、纯度、分裂国家的DNA在聚合酶链反应和抑制(PCR)。时间定性PCR方法检测35 s启动子和花椰菜花叶病毒根癌土壤杆菌tno在DNA序列样本提取认证参考材料和转基因大豆面粉样品了。最后,准备的标准曲线解释验证定量实时PCR分析达到理想值为-3.62lec参考基因和-3.40。农应变CP45-enolpyruvulshikimate-3-phosphate合酶(EPSPS)酶目标基因。量化的性能标准(within-laboratory再现性标准偏差、相对标准偏差不确定性、偏见、检测极限的极限量化和线性),因此本研究验证的方法。文件强调了一个明显的例子食品和饲料中转基因生物样本分析,指出需要多个实验室的研究在国家和国际水平。

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