Qualitative and quantitative polymerase chain reaction analysis for genetically modified maize MON863

Area of Science:

• Agricultural Biotechnology
• Molecular Biology
• Food Science

Background:

• Genetically modified (GM) crops require reliable detection methods.
• MON863 is a specific event of GM maize.
• Accurate qualitative and quantitative analysis is crucial for regulatory compliance and food safety.

Purpose of the Study:

• To develop and validate qualitative and quantitative analytical methods for GM maize MON863.
• To establish sensitive and specific polymerase chain reaction (PCR) assays for MON863 detection.

Main Methods:

• Qualitative detection using a specific primer pair for polymerase chain reaction (PCR).
• Quantitative detection using a specific primer pair and probe for real-time PCR.
• Development of a reference molecule incorporating maize DNA, CaMV 35S promoter, and MON863 construct-specific sequences.

Main Results:

• The qualitative PCR method demonstrated high specificity and sensitivity, with a limit of detection at 0.01% for GM maize MON863.
• The quantitative real-time PCR method showed a limit of quantitation of 0.5% for MON863.
• Validation at the 3.0% level yielded a bias of 3.0% and a relative standard deviation of 5.5%.

Conclusions:

• Developed PCR-based methods are effective for both qualitative and quantitative detection of GM maize MON863.
• These validated methods support the accurate identification and quantification of MON863 in food and agricultural products.
• The findings contribute to robust analytical tools for GM crop monitoring and regulation.