Establishment of Qualitative and Quantitative Detection Method for Transgenic Brassica napus NS-B50027-4
LEI Zhan1, WANG Jian-Cheng3, ZHANG Chen1, LI Kai2,3, HUANG Kun-Lun3, SHANG Ying1, XU Wen-Tao2,3
1 College of agriculture and food, Kunming University of Science and Technology, Kunming 650504, China;
2. Beijing Advanced Innovation Center for Food Nutrition and Human Health/Department of Nutrition and Health, China Agricultural University, Beijing 100083, China;
3 Key Laboratory of Safety Assessment of Genetically Modified Organism (Food Safety), Ministry of Agriculture / College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China
摘要转基因油菜(Brassica napus) NS-B50027-4是由澳大利亚-新西兰食品标准局批准的含有高比例的ω3脂肪酸和LC-ω3脂肪酸油菜新品种,到目前为止尚无文献报道对该转基因油菜新品种的检测方法,因此需要建立针对该品种的定性定量检测方法。本研究根据油菜内参基因HMG (high mobile group protein)和转基因油菜NS-B50027-4的A02染色体分别设计了引物和探针,利用普通PCR和实时荧光定量PCR技术分别测定引物和探针的特异性、灵敏度、准确性,从而确定该检测方法的检测限。实验结果表明,设计的两种引物都只能扩增出靶标,hmg引物的检测限能达到5 copies/μL,A02dn2引物的检测限达到1 copies/μL。因此,建立的转基因油菜NS-B50027-4的定性定量检测方法具有高度的特异性和选择性,优异的灵敏度和准确性。该研究结果为转基因油菜NS-B50027-4的定性定量检测提供了技术支持。
Abstract：Genetically modified Brassica napus NS-B50027-4 is a new variety containing a high proportion of ω3 fatty acids and LC-ω3 fatty acids approved by the Australia-New Zealand Food Standards Agency. So far there is no literature report on the detection method of this new genetically modified B. napus variety. Therefore, it is necessary to establish a qualitative and quantitative detection method for this species. In this study, primers and probes were designed according to the internal reference gene HMG (high mobile group protein) of B. napus and A02 chromosome of transgenic B. napus NS-B50027-4, respectively. The specificity, sensitivity and accuracy of the primers and probes were determined by conventional PCR and qPCR technology to determine the detection limit and quantification limit of the detection method. The experimental results show that the two designed primers could only amplify the target, the detection limit of hmg primer can reach 5 copies/μL, and the detection limit of A02dn2 primer is 1 copy/μL. Therefore, the established qualitative and quantitative detection method of genetically modified B. napus NS-B50027-4 has high specificity and selectivity, excellent sensitivity and accuracy. The research results provide technical support for the qualitative and quantitative detection of genetically modified B. napus NS-B50027-4.
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