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使用两种多重液滴数字PCR检测方法检测大豆转基因事件

Detection of Soybean GMO Events Using Two Multiplex Droplet Digital PCR Assays.

作者信息

Demeke Tigst, Eng Monika

机构信息

Canadian Grain Commission, Grain Research Laboratory, Winnipeg, MB R3C 3G8, Canada.

出版信息

J AOAC Int. 2025 Jan 1;108(1):23-28. doi: 10.1093/jaoacint/qsae082.

Abstract

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.

摘要

背景

由于法规合规要求,需要转基因生物事件的检测方法。高效检测和定量转基因生物事件可节省时间和资源。多重数字PCR(dPCR)可同时检测和定量多种转基因生物事件。

目的

本研究使用两种四重液滴数字PCR(ddPCR)检测方法来检测19种大豆转基因生物事件。

方法

开发并优化了两种多重dPCR检测方法,用于检测19种大豆转基因生物事件。第一种四重ddPCR检测方法包含转基因植物中常见的四个元件特异性靶标(P-35S、T-nos、tE9和Pat)。第二种事件特异性四重ddPCR检测方法针对四种不能被元件特异性四重ddPCR检测到的大豆转基因生物事件(CV127、DP305423、MON87701和MON87751)。

结果

元件特异性四重ddPCR检测方法检测到了所有预期的15种大豆转基因生物事件。元件特异性四重ddPCR检测方法还在0.01%的水平上检测到了选定的大豆转基因生物事件。事件特异性四重ddPCR检测方法成功地在0.1%、1%、2%和5%的水平上对四种大豆转基因生物事件进行了定量。事件特异性四重ddPCR检测方法也在0.01%的水平上检测到了这四种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种大豆转基因生物事件。

要点

使用元件特异性四重ddPCR检测方法检测15种大豆转基因生物事件,使用事件特异性四重ddPCR检测方法检测和定量四种不能被元件特异性ddPCR检测方法检测到的大豆转基因生物事件。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7b5a/11761968/05b3d467ad59/qsae082f1.jpg

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