ISO/TS 21569-7:2022 分子生物标志物分析的水平方法 转基因生物和衍生产品检测的分析方法 第7部分：CaMV和农杆菌Ti质粒衍生DNA序列检测的实时PCR方法

标准编号：ISO/TS 21569-7:2022

中文名称：分子生物标志物分析的水平方法 转基因生物和衍生产品检测的分析方法 第7部分：CaMV和农杆菌Ti质粒衍生DNA序列检测的实时PCR方法

英文名称：Horizontal methods for molecular biomarker analysis — Methods of analysis for the detection of genetically modified organisms and derived products — Part 7: Real-time PCR based methods for the detecti

发布日期：2022-12

标准范围

本文件规定了用于检测来自花椰菜花叶病毒(CaMV)的开放阅读框5(ORF V)的DNA序列的方法和用于检测来自植物病原性根瘤菌(以前称为根癌农杆菌)的肿瘤诱导(Ti)质粒的胭脂碱合酶(nos)基因的DNA序列的方法。所述方法可用于筛选遗传修饰的作物/植物及其衍生产物，以进一步阐明CaMV的特异性启动子或终止子(P-35S、T-35S)或两者以及nos基因(P-nos、T-nos)的阳性PCR结果。本文件中指定的方法将检测和鉴定天然存在的CaMV或放射根瘤菌(Ti质粒)DNA，或两者，如果在不存在含有指定靶序列的遗传修饰植物事件的情况下存在于样品中。这两种方法都基于实时聚合酶链反应（PCR），适用于从食品和其他产品（如饲料和种子/谷物）中提取的DNA的分析。这些方法的应用需要从相关基质中提取足够量的可扩增DNA。利用适当的校准材料，可以估计CaMV ORF V或nos拷贝数或两者，并分别与启动子(P-35S，P-nos)或终止子(T-35S，T-nos)序列或两者的估计拷贝数进行比较。因此，关于在测试样品中除了任何检测到的CaMV DNA或放射根瘤菌Ti质粒DNA或两者之外还存在未知的遗传修饰生物体(GMO)的结论是可能的。

This document specifies a procedure for the detection of a DNA sequence of the open reading frame five (ORF V) from cauliflower mosaic virus (CaMV) and a procedure for the detection of the DNA sequence of the nopaline synthase (nos) gene from tumour-inducing (Ti) plasmids of phytopathogenic Rhizobium radiobacter (formerly named Agrobacterium tumefaciens). The procedures can be used in the context of screening for genetically modified crop/plants and their derived products to further clarify a positive PCR result for a specific promoter or terminator of CaMV (P-35S, T-35S), or both, and the nos gene (P-nos, T-nos), respectively.

The methods specified in this document will detect and identify naturally occurring CaMV or Rhizobium radiobacter (Ti plasmid) DNA, or both, if present in the sample in the absence of a genetically modified plant event containing the specified target sequences.

Both methods are based on the real-time polymerase chain reaction (PCR) and are applicable for the analysis of DNA extracted from foodstuffs and other products such as feedstuffs and seeds/grains. The application of the methods requires the extraction of an adequate amount of amplifiable DNA from the relevant matrix.

With appropriate calibration material, the CaMV ORF V or nos copy number, or both, can be estimated and compared, respectively, with the estimated copy number for the promoter (P-35S, P-nos) or the terminator (T-35S, T-nos) sequences, or both. Thereby, conclusions are possible about the presence of an unknown genetically modified organism (GMO) in addition to any detected CaMV DNA or Rhizobium radiobacter Ti plasmid DNA, or both, in a test sample.

标准预览图