GMO Testing Laboratory, Experiment Research Institute of National Agricultural Products Quality Management Service, Seoul, South Korea.
J Agric Food Chem. 2009 Aug 26;57(16):7178-85. doi: 10.1021/jf901078d.
Analytical methods are very important in the control of genetically modified organism (GMO) labeling systems or living modified organism (LMO) management for biotech crops. Event-specific primers and probes were developed for qualitative and quantitative analysis for biotech maize event 3272 and LY 038 on the basis of the 3' flanking regions, respectively. The qualitative primers confirmed the specificity by a single PCR product and sensitivity to 0.05% as a limit of detection (LOD). Simplex and duplex quantitative methods were also developed using TaqMan real-time PCR. One synthetic plasmid was constructed from two taxon-specific DNA sequences of maize and two event-specific 3' flanking DNA sequences of event 3272 and LY 038 as reference molecules. In-house validation of the quantitative methods was performed using six levels of mixing samples, from 0.1 to 10.0%. As a result, the biases from the true value and the relative deviations were all within the range of +/-30%. Limits of quantitation (LOQs) of the quantitative methods were all 0.1% for simplex real-time PCRs of event 3272 and LY 038 and 0.5% for duplex real-time PCR of LY 038. This study reports that event-specific analytical methods were applicable for qualitative and quantitative analysis for biotech maize event 3272 and LY 038.
分析方法在转基因生物（GMO）标签系统的控制或生物技术作物的活体改性生物体（LMO）管理中非常重要。基于 3'侧翼区，分别为生物技术玉米事件 3272 和 LY 038 开发了事件特异性引物和探针进行定性和定量分析。定性引物通过单个 PCR 产物确认特异性，检测限（LOD）为 0.05%。还使用 TaqMan 实时 PCR 开发了 simplex 和 duplex 定量方法。从玉米的两个分类特异性 DNA 序列和事件 3272 和 LY 038 的两个事件特异性 3'侧翼 DNA 序列构建了一个合成质粒作为参考分子。使用 6 个混合水平（从 0.1 到 10.0%）对内定量方法进行了验证。结果，真实值的偏差和相对偏差均在 +/-30%范围内。定量方法的定量限（LOQ）对于事件 3272 和 LY 038 的 simplex 实时 PCR 均为 0.1%，对于 LY 038 的 duplex 实时 PCR 为 0.5%。本研究报告，事件特异性分析方法适用于生物技术玉米事件 3272 和 LY 038 的定性和定量分析。
