Abstract

This study focuses on developing a multiplex PCR assay for the simultaneous detection of four specific living modified (LM) maize varieties (DP-004114-3, MON89034, MON88017, and MON87403) approved for use in South Korea. The increasing use of LMO maize in food, feed, and industrial applications has heightened concerns about unintentional environmental releases, underscoring the need for reliable detection methods. Genetic information for each LMO event was obtained, and event-specific primers were designed to ensure accurate amplification. The developed multiplex PCR method underwent rigorous validation through specificity assays, analysis of certified reference material (CRM) mixtures, and limit of detection (LOD) testing. The assay effectively detected all target events with high sensitivity, capable of identifying as little as 12.5 ng μL–1 of genomic DNA. Furthermore, this method was utilized with suspicious samples gathered during the monitoring of unintentional LMO releases in natural settings, confirming its utility in practical applications. Our results show that the multiplex PCR assay is a robust and efficient method for the detection and monitoring of LM maize in both environmental and regulatory contexts. This technique holds considerable potential to safeguard natural ecosystems by preventing unintentional LMO releases and ensuring adherence to biosafety regulations.