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三种商业试剂盒与内部优化的聚合酶链式反应(PCR)检测方法在食品和饲料转基因生物筛查中的比较评估

Comparative assessment of three commercial kits and in house optimized PCR assays for GMO screening in food and feed.

作者信息

Verginelli Daniela, Quarchioni Cinzia, Spinella Katia, La Rocca Davide, Bonini Pamela, Fusco Cristiana, Misto Marisa, Peddis Stefania, Peroni Lorella, Marchesi Ugo

机构信息

National Reference Laboratory for GM Food and Feed, GMO Unit, Istituto Zooprofilattico Sperimentale del Lazio e della Toscana "Mariano Aleandri", Rome, Italy.

出版信息

MethodsX. 2024 Jul 26;13:102878. doi: 10.1016/j.mex.2024.102878. eCollection 2024 Dec.

Abstract

Screening strategies for GMO detection in food and feed are a crucial aspect in GMO testing laboratories for streamlining the analytical workflow and reducing turnaround time and costs. These strategies can be more or less complex or even be targeted according to the ingredients in the product, but whatever the choice, a good basic approach is generally based on the search for 35S promoter (P35S), nos-terminator (T-nos) and FMV promoter (P-FMV). In this study, we compare the singleplex real time PCR method for P35S, T-nos and P-FMV detection currently adopted by the Italian National Reference Laboratory for GM food and feed (NRL) with three commercial kits available on the market for giving a greater choice to consider the best approach suitable to the official control laboratories that are different from each other.•The NRL optimized singleplex PCR methods and the three commercial kits fully respect all the validation parameters criteria according to the minimum performance requirements (MPR) of ENGL [1]•Screening strategies for GMO detection in food and feed are a crucial aspect in GMO testing laboratories and being the commercial kits different from each other, the laboratory can choose the methods best suit their needs reducing turnaround time and costs.

摘要

食品和饲料中转基因生物(GMO)检测的筛选策略是转基因生物检测实验室的一个关键方面,有助于简化分析流程、减少周转时间和成本。这些策略可能或多或少较为复杂,甚至可根据产品中的成分进行针对性设计,但无论作何选择,一个好的基本方法通常是基于对35S启动子(P35S)、胭脂碱合成酶终止子(T-nos)和花椰菜花叶病毒35S启动子(P-FMV)的检测。在本研究中,我们将意大利国家转基因食品和饲料参考实验室(NRL)目前采用的用于检测P35S、T-nos和P-FMV的单重实时荧光定量PCR方法与市场上现有的三种商业试剂盒进行比较,以便提供更多选择,从而找出最适合不同官方检测实验室的最佳方法。•NRL优化的单重PCR方法和这三种商业试剂盒完全符合欧洲转基因生物检测实验室网络(ENGL)根据最低性能要求(MPR)制定的所有验证参数标准[1]•食品和饲料中转基因生物检测的筛选策略是转基因生物检测实验室的一个关键方面,鉴于商业试剂盒各不相同,实验室可以选择最适合自身需求的方法,以减少周转时间和成本。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/530f/11345693/74a3a809bd9f/ga1.jpg

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