A high-throughput screening method for GM soybean events based on single universal primer multiplex PCR and capillary electrophoresis

IF 1.4 3区 农林科学 Q4 FOOD SCIENCE & TECHNOLOGY Journal of Consumer Protection and Food Safety Pub Date : 2024-08-28 DOI:10.1007/s00003-024-01524-6
Liming Dong, Congcong Li, Zhenjuan Xing, Yuxuan He, Wei Xia, Yijun Liu, Likun Long, Feiwu Li
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Abstract

Genetically modified (GM) crops and their products are regulated in many countries. To meet increasing regulatory requirements for a growing number of GM soybean events, high-throughput and cost-effective detection methods are indispensable. In this study, 14 widely commercialized GM soybean events, including GTS40-3-2, MON89788, CV127, A2704-12, A5547-127, 305423, 356043, MON87701, MON87705, MON87708, MON87769, SYHT0H2, FG72, and DAS-444Ø6-6 were utilized as targets for event-specific identification. A high-throughput detection method was established by integrating a single universal primer multiplex PCR (SUP-M-PCR) with capillary electrophoresis technique. This method enables the identification of 14 soybean events in a single PCR reaction, addressing the limitations of conventional multiplex PCR such as self-inhibition and amplification variations between different primers. Furthermore, it significantly enhances the sensitivity and accuracy of previous multiple detection methods, achieving a detection sensitivity of 0.05% (w/w). Overall, this approach greatly improves detection efficiency and holds promising potential for the identification and detection of 14 types of GM soybean events and their derivatives.

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基于单一通用引物多重 PCR 和毛细管电泳的转基因大豆事件高通量筛选方法
转基因作物及其产品在许多国家都受到监管。为了满足日益增多的转基因大豆事件的监管要求,高通量、高性价比的检测方法必不可少。本研究以 GTS40-3-2、MON89788、CV127、A2704-12、A5547-127、305423、356043、MON87701、MON87705、MON87708、MON87769、SYHT0H2、FG72 和 DAS-444Ø6-6 等 14 个广泛商业化的转基因大豆事件为目标,进行事件特异性鉴定。通过将单一通用引物多重 PCR(SUP-M-PCR)与毛细管电泳技术相结合,建立了一种高通量检测方法。该方法可在一次 PCR 反应中鉴定 14 个大豆事件,解决了传统多重 PCR 的局限性,如自抑制和不同引物之间的扩增差异。此外,该方法还大大提高了以往多重检测方法的灵敏度和准确性,检测灵敏度达到 0.05%(重量比)。总之,这种方法大大提高了检测效率,有望用于 14 种转基因大豆事件及其衍生物的鉴定和检测。
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来源期刊
CiteScore
3.70
自引率
4.20%
发文量
46
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.
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