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“组合 SYBRGreen qPCR 筛选”的理论介绍,一种基于矩阵的方法,用于检测来自转基因植物的材料。

A theoretical introduction to "combinatory SYBRGreen qPCR screening", a matrix-based approach for the detection of materials derived from genetically modified plants.

机构信息

Division of Biosafety and Biotechnology, Scientific Institute of Public Health, J. Wytsmanstreet 14, 1050 Brussels, Belgium.

出版信息

Anal Bioanal Chem. 2010 Mar;396(6):2113-23. doi: 10.1007/s00216-009-3286-7. Epub 2009 Dec 4.

Abstract

The detection of genetically modified (GM) materials in food and feed products is a complex multi-step analytical process invoking screening, identification, and often quantification of the genetically modified organisms (GMO) present in a sample. "Combinatory qPCR SYBRGreen screening" (CoSYPS) is a matrix-based approach for determining the presence of GM plant materials in products. The CoSYPS decision-support system (DSS) interprets the analytical results of SYBRGREEN qPCR analysis based on four values: the C(t)- and T(m) values and the LOD and LOQ for each method. A theoretical explanation of the different concepts applied in CoSYPS analysis is given (GMO Universe, "Prime number tracing", matrix/combinatory approach) and documented using the RoundUp Ready soy GTS40-3-2 as an example. By applying a limited set of SYBRGREEN qPCR methods and through application of a newly developed "prime number"-based algorithm, the nature of subsets of corresponding GMO in a sample can be determined. Together, these analyses provide guidance for semi-quantitative estimation of GMO presence in a food and feed product.

摘要

食品和饲料产品中转基因(GM)物质的检测是一个复杂的多步骤分析过程,需要对样品中存在的转基因生物(GMO)进行筛选、鉴定,并且通常还需要进行定量分析。“组合 qPCR SYBRGreen 筛选”(CoSYPS)是一种基于矩阵的方法,用于确定产品中 GM 植物材料的存在。CoSYPS 决策支持系统(DSS)根据四个值(每个方法的 C(t)值和 T(m)值以及 LOD 和 LOQ)解释 SYBRGREEN qPCR 分析的分析结果。本文给出了 CoSYPS 分析中应用的不同概念的理论解释(GMO 宇宙、“素数追踪”、矩阵/组合方法),并以 RoundUp Ready 大豆 GTS40-3-2 为例进行了说明。通过应用有限数量的 SYBRGREEN qPCR 方法,并应用新开发的“素数”算法,可以确定样品中相应 GMO 的子集的性质。这些分析共同为食品和饲料产品中转基因物质存在的半定量估计提供了指导。

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