转基因大豆DAS44406-6多重荧光定量PCR检测方法的建立

发布时间：2018-10-09 09:04

【摘要】：本文针对大豆内源基因Lectin和转基因大豆DAS44406-6品系的5'端插入位点序列,设计特异性引物及探针,建立同时检测转基因大豆DAS44406-6品系和大豆内源基因Lectin的多重荧光定量PCR方法,并运用15种转基因大豆、3种转基因玉米、1种转基因油菜、1种转基因水稻和非转基因大豆对该方法进行特异性评价,并分析该方法的灵敏度和稳定性。结果显示,该方法能准确从20种转基因样品和1种非转基因样品中检出靶目标,检测结果与待检样品信息一致,表明本方法具有良好的特异性;灵敏度高达0.01%;重复性实验表明DAS81419品系4种含量、9次重复反应Ct值的标准偏差介于0.050~0.222,相对标准偏差介于0.169%~0.677%,均在可接受范围内。该方法特异性强、灵敏度高、稳定性强,适用于各口岸实验室进行转基因大豆DAS44406-6的快速、准确的检测。
[Abstract]:In this paper, specific primers and probes were designed to detect the 5 'terminal insertion site sequences of soybean endogenous gene Lectin and transgenic soybean DAS44406-6 strain, and a multiplex fluorescence quantitative PCR method was established for simultaneous detection of DAS44406-6 strain and Lectin gene in transgenic soybean. The sensitivity and stability of this method were evaluated by using 15 kinds of transgenic soybean, 3 kinds of transgenic corn, 1 kind of transgenic rape and 1 kind of transgenic rice and non-transgenic soybean, and the sensitivity and stability of the method were analyzed. The results showed that the method could detect target accurately from 20 kinds of transgenic samples and 1 kind of non-transgenic samples. The detection results were consistent with the information of the samples to be tested, which indicated that the method had good specificity. The reproducibility test showed that the standard deviation of Ct value of the four DAS81419 strains was between 0.050 and 0.222, and the relative standard deviation was between 0.169 and 0.677, which was within the acceptable range. This method has the advantages of high specificity, high sensitivity and strong stability. It is suitable for rapid and accurate detection of transgenic soybean DAS44406-6 in port laboratories.
【作者单位】：中国检验检疫科学研究院;汕头出入境检验检疫局;湛江出入境检验检疫局;暨南大学理工学院食品科学与工程系;
【基金】：广东省科技计划项目(2014A040401029) 转基因产品抽制样和精准检测技术(2016ZX08012-001) 广州市科技计划项目(2014J4100105)
【分类号】：S565.1

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