Event‐specific analytical methods for biotech maize MIR 604 and DAS‐59122‐7

BACKGROUND: It is very important to develop analytical methods for genetically modified organism (GMO) labelling systems and living modified organism (LMO) management. The polymerase chain reaction (PCR) is the most efficient DNA‐based analytical method for identifying and quantifying biotech crops. Qualitative PCR methods have been developed to detect the presence of biotech crops, while quantitative PCR methods have been developed to analyse the content of biotech crops. Analytical methods are now required for new biotech maize events, MIR 604 and DAS‐59122‐7. RESULTS: The event‐specific primers and probes were developed for qualitative and quantitative analysis of biotech maize MIR 604 and DAS‐59122‐7 based on the 3′ flanking regions. As a reference molecule, single standard plasmid was developed. The specificity of the qualitative primers was confirmed by the single PCR product, with limits of detection (sensitivity) of 0.01 and 0.05% respectively. In‐house validation of the quantitative methods was performed using six levels of mixing samples (0.1–10% w/w). As a result, the biases from the true value and the relative standard deviations were within the range of ± 30%. The limits of quantitation of the quantitative methods were all 0.1% for real time PCRs of MIR 604 and DAS‐59122‐7. CONCLUSION: In this study, event‐specific analytical methods were developed and applied to the qualitative and quantitative analysis of biotech maize MIR 604 and DAS‐59122‐7. Copyright © 2009 Society of Chemical Industry