Development and validation of a multiplex real-time PCR method for screening genetically modified plants

IF 1.4 3区农林科学 Q4 FOOD SCIENCE & TECHNOLOGY

Journal of Consumer Protection and Food Safety Pub Date : 2024-02-24 DOI:10.1007/s00003-024-01499-4

Christopher Weidner, René Köppel, Regina Freyer, Petra Richl, Kathrin Lieske, Joachim Mankertz, Hans-Ulrich Waiblinger

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引用次数: 0

Abstract

The diversity of genetically modified plants used for food and feed is increasing worldwide. For the detection and control of these products, efficient and reliable analytical tools are a prerequisite. This can be done by screening for specific DNA-elements and constructs characteristic of transgenic plants. In the past, numerous methods have already been published. However, several genetically modified plants are not covered by common screening methods. Here, a new qualitative triplex real-time polymerase chain reaction (PCR) method is presented, detecting two transgene flanking sequences and the transition between the Cassava Vein Mosaic Virus Promotor (P-CsVMV) and the phosphinothricin-N-acetyltransferase (pat) gene. These sequences are present in several transgenic plants and therefore, the described triplex method can be used as a screening tool to guide further analysis and increase the efficiency of the analysis strategy for GMO detection. The method is characterized by high specificity, sensitivity and robustness and is provided as a ring-trial validated method in the Official Collection of Methods according to the German Food and Feed Act.

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开发和验证用于筛选转基因植物的多重实时 PCR 方法

全球范围内用于食品和饲料的转基因植物日益多样化。要检测和控制这些产品，高效可靠的分析工具是先决条件。这可以通过筛选转基因植物特有的 DNA 元素和构建体来实现。过去，已经有许多方法发表。然而，一些转基因植物并不在常用筛选方法的范围内。本文介绍了一种新的定性三重实时聚合酶链反应（PCR）方法，该方法可检测两个转基因侧翼序列以及木薯花叶病毒启动子（P-CsVMV）和膦丝菌素-N-乙酰转移酶（pat）基因之间的过渡。这些序列存在于几种转基因植物中，因此，所述三重法可用作筛选工具，指导进一步的分析，并提高转基因生物检测分析策略的效率。该方法具有特异性高、灵敏度高和稳健性强的特点，已作为环试验证方法列入《德国食品和饲料法官方方法集》。

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来源期刊

Journal of Consumer Protection and Food Safety Veterinary-Food Animals

CiteScore

3.70

自引率

4.20%

发文量

审稿时长

>12 weeks

期刊介绍： The JCF publishes peer-reviewed original Research Articles and Opinions that are of direct importance to Food and Feed Safety. This includes Food Packaging, Consumer Products as well as Plant Protection Products, Food Microbiology, Veterinary Drugs, Animal Welfare and Genetic Engineering. All peer-reviewed articles that are published should be devoted to improve Consumer Health Protection. Reviews and discussions are welcomed that address legal and/or regulatory decisions with respect to risk assessment and management of Food and Feed Safety issues on a scientific basis. It addresses an international readership of scientists, risk assessors and managers, and other professionals active in the field of Food and Feed Safety and Consumer Health Protection. Manuscripts – preferably written in English but also in German – are published as Research Articles, Reviews, Methods and Short Communications and should cover aspects including, but not limited to: · Factors influencing Food and Feed Safety · Factors influencing Consumer Health Protection · Factors influencing Consumer Behavior · Exposure science related to Risk Assessment and Risk Management · Regulatory aspects related to Food and Feed Safety, Food Packaging, Consumer Products, Plant Protection Products, Food Microbiology, Veterinary Drugs, Animal Welfare and Genetic Engineering · Analytical methods and method validation related to food control and food processing. The JCF also presents important News, as well as Announcements and Reports about administrative surveillance.