Quantitative detection method for Roundup Ready soybean in food using duplex real-time PCR MGB chemistry

Area of Science:

• Food safety and biotechnology
• Molecular biology and genetic engineering

Background:

• Accurate detection of genetically modified organisms (GMOs) is crucial for legislation and labeling.
• Increasing numbers of approved genetically modified (GM) lines necessitate cost-efficient detection methods.
• Quantitative polymerase chain reaction (qPCR) offers sensitivity and specificity but can be costly.

Purpose of the Study:

• To develop a multiplexing approach for cost-efficient GMO detection.
• To create a duplex qPCR assay for specific quantification of Roundup Ready soybean (RRS) event GTS 40-3-2.

Main Methods:

• Development of construct-specific primers and probes for RRS event GTS 40-3-2.
• Utilization of the lectin gene (Le1) as a specific reference gene.
• Optimization and validation of a duplex quantitative real-time PCR (qRTPCR) platform using TaqMan MGB-NFQ chemistry.

Main Results:

• A duplex qRTPCR assay was successfully developed for RRS event GTS 40-3-2 detection.
• The assay demonstrated validated limits of detection (LOD) and quantification (LOQ), along with accuracy.
• DNA degradation in processed food samples had minimal adverse effects on the assay's performance.

Conclusions:

• A duplex qRTPCR assay was established for the specific detection and quantification of RRS event GTS 40-3-2.
• The developed method is suitable for practical monitoring of this GM event in processed foods.
• This approach offers a cost-effective solution for GMO testing in food products.