Electrochemical detection of 0.6% genetically modified maize MON810 in real flour samples
Electrochemical detection of 0.6% genetically modified maize MON810 in real flour samples
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DOI：
10.1016/j.elecom.2012.06.019
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发表时间：
2012-08
影响因子：
5.4
通讯作者：
Maren Mix;Julia Rüger;S. Krüger;I. Broer;G. Flechsig
中科院分区：
文献类型：
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作者：
Maren Mix;Julia Rüger;S. Krüger;I. Broer;G. Flechsig
我们展示了一种方法，通过方波伏安法（SWV）的转基因玉米在真实的玉米粉样品的电化学检测。用四氧化锇联吡啶[OsO 4（bipy）]标记不对称PCR扩增的靶标后，它们与固定在金电极上的寡核苷酸探针杂交。在近等基因玉米中检测到玉米基因ivrp和SSIIb。在所有转基因玉米样品中均检测到转基因cryIa/B和MON 810特异性片段，在混合样品中MON 810的含量低至0.6%。虽然在少于10分钟的杂交时间后可以检测到含有100%近等基因或相应的转基因玉米的样品中的所有序列，但是对于检测仅含有0.9%至0.6%的转基因玉米的样品中的遗传修饰，需要30分钟的杂交时间。当样品中仅存在0.5%的转基因玉米时，不可能显著检测到转基因cryIa/B或MON 810，这很可能是由于模板DNA的扩增不足。
We demonstrate an approach for the electrochemical detection of genetically modified maize in real maize flour samples by means of square-wave voltammetry (SWV). After labeling the asymmetric PCR-amplified targets with osmium tetroxide bipyridine [OsO4(bipy)], they were hybridized with immobilized oligonucleotide probes on gold electrodes. We could detect the maize genes ivrp and SSIIb in near isogenic maize. The transgene cryIa/b and the MON810 specific fragment were detected in all transgenic maize samples, down to a content of 0.6% of MON810 in mixed samples. While it was possible to detect all sequences in the samples containing 100% near isogenic or respectively transgenic maize after a hybridization time of less than 10min, a hybridization time of 30min was necessary for the detection of the genetic modifications in samples containing only 0.9 to 0.6% of transgenic maize. No significant detection of the transgene cryIa/b or MON810 was possible when only 0.5% of transgenic maize was present in the sample, most likely due to insufficient amplification of the template DNA.