Development of quantitative duplex real-time PCR method for screening analysis of genetically modified maize

Area of Science:

• Agricultural Biotechnology
• Molecular Biology
• Analytical Chemistry

Background:

• Quantitative analysis of genetically modified organisms (GMOs) in food is crucial for regulatory compliance and consumer safety.
• Existing simplex real-time PCR methods, while standardized, can be time-consuming and costly for routine screening.
• The need for efficient and precise methods for detecting specific genetically modified maize events is ongoing.

Purpose of the Study:

• To develop and evaluate a duplex real-time PCR method for the quantitative screening of genetically modified maize.
• To simultaneously detect the cauliflower mosaic virus (CaMV) 35S promoter (P35S) and an event-specific segment for GA21 maize.
• To assess the analytical performance and efficiency of the developed duplex method compared to existing techniques.

Main Methods:

• Development of a duplex real-time PCR assay targeting two distinct DNA sequences in GM maize.
• Utilization of a specifically designed plasmid calibrant for accurate quantification.
• In-house evaluation of the method's analytical precision and efficiency.

Main Results:

• The duplex real-time PCR method demonstrated analytical precision comparable to established simplex methods.
• The assay successfully detected both the CaMV 35S promoter (P35S) and an event-specific segment for GA21 maize simultaneously.
• The developed method has the potential to reduce the cost and time for routine GMO analysis by approximately 50%.

Conclusions:

• The developed duplex real-time PCR method is a high-performing tool for the quantitative screening of GM maize.
• This method offers significant advantages in terms of cost and time efficiency for routine GMO analysis.
• Further interlaboratory collaborative studies are recommended to validate the method for broader practical application.