Chinese Academy of Inspection and Quarantine, Room 614, Building 241, HUIXINLI, Huixinxi Street, Chaoyang District, Beijing 100029, China.
Biosens Bioelectron. 2013 Nov 15;49:360-6. doi: 10.1016/j.bios.2013.05.045. Epub 2013 Jun 6.
An integration event-specific fluorescent liquid bead array was developed for the simultaneous identification of 10 genetically modified (GM) maize, including Bt176, Bt11, MON810, NK603, GA21, MON88017, MON89034, MIR604, T25 and MIR162, as well as one non-GM maize. The system comprised 11 specific oligonucleotide probes labeled with an amino group and coupled to fluorescence-encoded microspheres. To enable fluorescence detection, 11 pairs of primers labeled with biotin at the 5' ends were used. The hybridization signal of biotinylated PCR product to the probe-coupled microspheres was then detected. The limit of detection of this assay was 0.1% for GM maize, which is lower than the current labeling threshold levels enforced in the EU (0.9%). The results of the positive and negative controls were consistent with their expected situation, which showed that the method was highly specific. We detected GM maize in 20 of the 1370 commercial food samples tested, which were labeled as containing maize. The overall sensitivity, specificity, rapidity and high throughput capacity of this liquid chip system suggest that it could provide a significant improvement over current methods, and potentially offer an improved platform for further research into the detection of other GM plants.
建立了一种基于荧光液芯芯片的整合事件特异性检测方法，用于同时鉴定 10 种转基因玉米，包括 Bt176、Bt11、MON810、NK603、GA21、MON88017、MON89034、MIR604、T25 和 MIR162，以及一种非转基因玉米。该系统由 11 个带有氨基的特异性寡核苷酸探针和与荧光编码微球偶联组成。为了进行荧光检测，使用了 11 对带有生物素标记的 5' 端引物。然后检测生物素化 PCR 产物与探针偶联微球的杂交信号。该检测方法的检测限为转基因玉米 0.1%，低于欧盟现行的标记阈值（0.9%）。阳性和阴性对照的结果与预期一致，表明该方法具有高度特异性。我们在 1370 个被标记为含有玉米的商业食品样品中检测到了 20 个转基因玉米。该液芯芯片系统具有高灵敏度、特异性、快速和高通量的特点，有望优于现有方法，并为进一步研究其他转基因植物的检测提供一个改进的平台。
