转基因甜菜品系H7-1的数字PCR定量检测方法

发布时间：2018-04-16 15:17

  本文选题：转基因甜菜 + H-品系　； 参考：《现代食品科技》2017年07期

【摘要】：为实现转基因甜菜品系H7-1的标识管理和精准定量,根据H7-1的5’边界序列和甜菜谷氨酰胺合成酶基因(glutamine synthetase,GS)设计引物和探针建立双重数字PCR检测体系。该方法的特异性、灵敏度、精密度和准确度均进行了测试。结果显示:建立的转基因甜菜H7-1数字PCR检测方法特异于H7-1品系检测,在20μL反应体中H7-1品系特异性序列和内源基因GS的定量下限(limit of quantitation,LOQ)分别为3.1拷贝/μL和6.3拷贝/μL,检测下限(limit of detection,LOD)检出限分别为0.6拷贝/μL和1.3拷贝/μL,精密度和准确度在可接受范围内,该定量方法不依赖于标准曲线建立,可便捷的应用于转基因甜菜H7-1成分的精确定量检测。
[Abstract]:In order to achieve the label management and accurate quantification of transgenic sugarbeet strain H7-1, a double digital PCR detection system was established by designing primers and probes based on the 5'boundary sequence of H7-1 and the sugar beet glutamine synthetase gene (GS-GS).The specificity, sensitivity, precision and accuracy of the method were tested.The results showed that the established H7-1 digital PCR detection method was specific to H7-1 strain detection.In the 20 渭 L reaction, the limit of limit of quantification of the specific sequence and the endogenous gene GS were 3.1 copies / 渭 L and 6.3 copies / 渭 L, respectively, and the detection limit of detection limit of detection LOD was 0.6 copies / 渭 L and 1.3 copies / 渭 L, respectively, and the precision and accuracy were within the acceptable range.The method does not depend on the standard curve and can be applied to the accurate quantitative detection of H7-1 components in transgenic beet.
【作者单位】： 黄埔出入境检验检疫局;广东检验检疫技术中心广东省动植物与食品进出口技术措施研究重点实验室;
【基金】：广东省科技计划项目(2014A040401029) 出入境检验检疫行业标准计划项目(2015B218k) 广东出入境检验检疫局科技计划项目(2015GDK13)
【分类号】：TS255.7
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本文编号：1759487