A Multiplex PCR System for the Detection and Quantification of Four Genetically Modified Soybean Events

To align with global regulatory standards for quantitative GMO labeling, there is a growing need for accurate and efficient detection methodologies targeting on specific genetically modified (GM) events. This study developed a multiplex detection strategy for four GM soybean events (ZH10-6, ZUTS-33, SHZD32, and S4003.14), along with the endogenous Lectin gene. A 5-plex qPCR assay was optimized with event-specific primers and probes, enabling the reliable detection of these GM events in mixtures at concentrations as low as 0.1%. Furthermore, two 3-plex cdPCR systems were established on the Naica chip-based platform, achieving limits of detection (LOD) and quantification (LOQ) at 10 and 20 copies, respectively, for each target event. Validation using test samples demonstrated that the accuracy and precision of these methods meet regulatory requirements for GMO labeling. Cross-platform comparisons between the digital PCR systems showed consistent results, improving both detection throughput and operational efficiency. This multiplex approach provides a robust and efficient solution for the quantitative analysis of GM soybean components in complex sample matrices, offering valuable technical support for regulatory compliance and GMO product monitoring.