本文将可视化DNA微阵列系统与多重PCR技术耦合来快速检测十二种转基因玉米。为了保证该方法的可靠性，本文在DNA微阵列中采用了正、反杂交控制。作者通过所研发的方法、PCR方法和测序方法对商业化的转基因材料（GMM：Bt176，Bt11，MON810，GA21、T25、MON88017，NK603，MON863，MON89034，：DAS-59122-7、TC1507，MIR604）；转基因棉花（MON1445, MON15985）；转基因大豆（孟山都公司抗农达大豆40-3-2）和非转基因材料进行检测。研究发现，该方法具有检测快速、成本效益高和易用性等优势。高特异性和敏感性的结果表明，可利用该方法进行转基因作物的常规分析。
外文摘要：
We herein developed a visual DNA microarray system coupled with multiplex PCR (m-PCR) to rapidly detect twelve genetically modified maize (GMM). The microarray comprised short oligonucleotide probes complimentary to the specific gene region for twelve different GMM. The m-PCR products annealed to the microarray probe were reacted with streptavidin-alkaline phosphatase conjugate and nitro blue tetrazolium/5-bromo-4-chloro-3'-indolylphosphate, p-toluidine salt (NBT/BCIP), resulting in blue spots that are easily visualized by unaided eyes for qualitative analysis. To ensure the reliability of this method, positive and negative hybridization controls were used in DNA microarray. Commercial GM materials (GMM: Bt176, Bt11, MON810, GA21, T25, MON88017, NK603, MON863, MON89034, DAS-59122-7, TC1507, MIR604); GM cotton: (MON1445, MON15985); GM soybean (Monsanto Roundup Ready soybean 40-3-2)) and non-GM materials were identified by this method and further confirmed by PCR and sequencing. The results showed that each probe consistently identified its corresponding GMM target very quickly and in a cost-effective and more time efficient way. The limit,of detection is 0.5% for Bt176, Bt11, T25, MON88017, DAS59122-7, MON89034 and 1% for MON810, MIR604, GA21, MON863, NK603, TC1507. This method is advantageous because of rapid detection, cost-effectiveness and ease of use. These high specificity and sensitivity results demonstrate the feasibility of using this method in routine analysis GMOs.
外文关键词：Visual detection；Multiplex PCR；Genetically modified maize；Microarray；Biochip
作者：Li, Yongjin；Xiong, Tao； Wu, Huawei；等
作者单位：Huzhou University
期刊名称：BIOCHIP JOURNAL
期刊影响因子：1.211
出版年份：2016
出版刊次：3
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