Interlaboratory validation of quantitative duplex real-time PCR method for screening analysis of genetically modified...

Area of Science:

• Food Science
• Molecular Biology
• Biotechnology

Background:

• Routine genetically modified organism (GMO) testing is time-consuming and costly.
• Accurate detection methods are crucial for food safety and regulatory compliance.

Purpose of the Study:

• To develop and validate a cost-effective and time-efficient duplex quantitative real-time PCR method for GMO screening.
• To simultaneously target event-specific segments for GA21 and the Cauliflower Mosaic Virus 35S promoter (P35S) in maize.

Main Methods:

• Development of a duplex quantitative real-time PCR assay.
• Determination of conversion factors (CFs) for two real-time PCR instruments (ABI PRISM 7900HT and 7500).
• Conducting an interlaboratory collaborative study including a blind test.

Main Results:

• The method achieved a limit of quantitation of 0.5% or less for both GA21 and P35S.
• Trueness and precision were evaluated, with determined bias and relative standard deviation of reproducibility (RSD(R)) each below 25%.
• The method demonstrated reliability across different instruments and laboratories.

Conclusions:

• The developed duplex quantitative real-time PCR method is suitable for practical screening analysis of GM maize.
• This method offers a reduced-cost and time-efficient alternative for routine GMO testing.
• The interlaboratory study confirmed the method's validity and reliability for screening GM maize.