使用两种多重液滴数字 PCR 检测大豆转基因生物事件。

Tigst Demeke, Monika Eng
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引用次数: 0

摘要

背景:由于监管合规要求,需要转基因生物事件的检测方法。对转基因生物事件进行高效检测和定量可节省时间和资源。多重数字 PCR(dPCR)可同时检测和定量多个转基因生物事件:本研究使用两种四联液滴数字 PCR(ddPCR)测定法检测了 19 个大豆转基因生物事件:方法:为检测 19 个大豆转基因生物事件,开发并优化了两种多重 dPCR 检测方法。第一种四重 ddPCR 检测包含转基因植物中常见的四种特异性靶标(P-35S、T-nos、tE9 和 Pat)。第二种事件特异性四重 ddPCR 检测针对的是元素特异性四重 ddPCR 检测不到的四种大豆转基因生物事件(CV127、DP305423、MON87701 和 MON87751):结果:元素特异性四聚体 ddPCR 检测法检测到了所有预期的 15 个大豆转基因生物事件。元素特异性四重 ddPCR 检测法还检测到了 0.01% 水平的部分大豆转基因生物事件。事件特异性四聚体 ddPCR 检测法成功地在 0.1%、1%、2% 和 5%的水平上对 4 个大豆转基因生物事件进行了量化。事件特异性四重 ddPCR 法还检测出了 0.01% 水平的四个大豆转基因生物事件:结论:这两种四重 ddPCR 检测方法可用于检测 19 个大豆转基因生物事件:使用元素特异性四联ddPCR检测法检测了15个大豆转基因生物事件,使用事件特异性四联ddPCR检测法检测并量化了元素特异性ddPCR检测法检测不到的4个大豆转基因生物事件。
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Detection of Soybean GMO Events Using Two Multiplex Droplet Digital PCR Assays.

Background: Detection methods for GMO events are required because of regulatory compliance requirements. Efficient detection and quantification of GMO events saves time and resources. Multiplex digital PCR (dPCR) allows detection and quantification of more than one GMO event at the same time.

Objective: The study used two tetraplex droplet digital PCR (ddPCR) assays for the detection of 19 soybean GMO events.

Methods: Two multiplex dPCR assays were developed and optimized for the detection of 19 soybean GMO events. The first tetraplex ddPCR assay contained four element-specific targets commonly found in GMO plants (P-35S, T-nos, tE9, and Pat). The second event-specific tetraplex ddPCR assay targeted four soybean GMO events that are not detected with the element-specific tetraplex ddPCR (CV127, DP305423, MON87701, and MON87751).

Results: The element-specific tetraplex ddPCR assay detected all the expected 15 soybean GMO events. The element-specific tetraplex ddPCR assay also detected selected soybean GMO events at the 0.01% level. The event-specific tetraplex ddPCR assay was successfully used to quantify the four soybean GMO events at the 0.1, 1, 2, and 5% levels. The event-specific tetraplex ddPCR assay also detected the four soybean GMO events at the 0.01% level.

Conclusions: The two tetraplex ddPCR assays can be used for the detection of 19 soybean GMO events.

Highlights: An element-specific tetraplex ddPCR assay was used to detect 15 soybean GMO events, and an event-specific tetraplex ddPCR assay was used to detect and quantify four soybean GMO events that are not detected by the element-specific ddPCR assay.

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