Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100176, China.
Shantou Entry-exit Inspection and Quarantine Bureau, Shantou 515041, China.
Food Chem. 2017 Aug 15;229:396-402. doi: 10.1016/j.foodchem.2017.02.088. Epub 2017 Feb 21.
High throughput screening systems are the preferred solution to meet the urgent requirement of increasing number of genetically modified organisms (GMOs). In this study, we have successfully developed a multiplex GMO element screening system with dual priming oligonucleotide (DPO) primers. This system can detect the cauliflower mosaic virus 35S (CaMV 35S), terminator of nopaline synthase gene (NOS), figwort mosaic virus 35S (FMV 35S) promoter, neomycin phosphotransferaseII (NPTII), Bt Cry 1Ab, phosphinothricin acetyltransferase genes (bar) and Streptomyces viridochromogenes (pat) simultaneously, which covers more than 90% of all authorized GMO species worldwide. This system exhibits a high tolerance to annealing temperatures, high specificity and a limit of detection equal to conventional PCR. A total of 214 samples from markets, national entry-exit agencies, the Institute for Reference Materials and Measurement (IRMM) and the American Oil Chemists' Society (AOCS) were also tested for applicability. This screening system is therefore suitable for GMO screening.
高通量筛选系统是满足不断增加的转基因生物（GMO）数量的迫切需求的首选解决方案。在本研究中，我们成功开发了一种使用双重引物寡核苷酸（DPO）引物的多重 GMO 元件筛选系统。该系统可以同时检测花椰菜花叶病毒 35S（CaMV 35S）、胭脂碱合成酶基因终止子（NOS）、首蓿花叶病毒 35S（FMV 35S）启动子、新霉素磷酸转移酶 II（NPTII）、Bt Cry 1Ab、膦丝菌素乙酰转移酶基因（bar）和变绿链霉菌（pat），涵盖了全球 90%以上的所有授权 GMO 物种。该系统对退火温度具有很高的耐受性、高度特异性和与常规 PCR 相当的检测限。还对来自市场、国家出入境机构、参考材料和测量研究所（IRMM）和美国油化学家协会（AOCS）的 214 个样本进行了适用性测试。因此，该筛选系统适用于 GMO 筛选。
