Development of a Multiplex Polymerase Chain Reaction–Capillary Gel Electrophoresis Assay for Detecting Seven Living Modified Canola Events
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Modern biotechnology has enabled significant improvements in crop production; however, the potential risks to human health and biodiversity remain concerning. Therefore, accurate and convenient detection methods for living modified organisms must be established to manage unintentional releases and increase the number of volunteers in natural ecosystems. This study successfully developed and validated two sets of multiplex polymerase chain reaction (PCR) methods for seven living modified organism canola events (Rf3, DP-073496-4, Topas19/2, Ms8, MON88302, GT73, and T45) and an endogenous gene (CruA) approved in South Korea. Capillary electrophoresis was used to analyze seven living modified organism canola events by combining multiplex PCR with fluorescence capillary electrophoresis, a technique commonly used in species identification and disease diagnosis. These newly developed multiplex PCR methods and capillary electrophoresis techniques were highly effective for analyzing living modified organism canola. Eight suspected living modified organism canola samples collected from ecosystems in South Korea were analyzed for practical use. These results suggest that the newly developed multiplex PCR and capillary electrophoresis techniques represent the first step toward automating the identification of living modified organisms.
Abstract
Modern biotechnology has enabled significant improvements in crop production; however, the potential risks to human health and biodiversity remain concerning. Therefore, accurate and convenient detection methods for living modified organisms must be established to manage unintentional releases and increase the number of volunteers in natural ecosystems. This study successfully developed and validated two sets of multiplex polymerase chain reaction (PCR) methods for seven living modified organism canola events (Rf3, DP-073496-4, Topas19/2, Ms8, MON88302, GT73, and T45) and an endogenous gene (CruA) approved in South Korea. Capillary electrophoresis was used to analyze seven living modified organism canola events by combining multiplex PCR with fluorescence capillary electrophoresis, a technique commonly used in species identification and disease diagnosis. These newly developed multiplex PCR methods and capillary electrophoresis techniques were highly effective for analyzing living modified organism canola. Eight suspected living modified organism canola samples collected from ecosystems in South Korea were analyzed for practical use. These results suggest that the newly developed multiplex PCR and capillary electrophoresis techniques represent the first step toward automating the identification of living modified organisms.