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发明公开
CN105112536A 转基因玉米NK603双重数字PCR荧光定量检测方法 无效 - 驳回

转基因玉米NK603双重数字PCR荧光定量检测方法
摘要：
本发明提供一种针对转基因玉米NK603的双重数字PCR荧光定量检测方法，本方法可以直接对待检测样品的基因组中的转基因玉米NK603进行定量检测，从而省去了常规PCR方法定量检测中标准曲线绘制步骤以及现有数字PCR检测中的样品前处理步骤，另外，在本方法中，由于转基因成分是通过外源基因拷贝数与内参基因拷贝数的比例计算得到的，因此在同一个反应体系进行双重检测可以提高转基因成分定量检测的稳定性。利用本方法可实现对转基因玉米NK603的绝对定量检测，定量检测限可达到0.5％，灵敏度可达到0.1％，能够满足所有转基因成分实际检测的需要。此外，本方法操作简单，灵活性强，是一种转基因绝对定量检测的有效方法。
摘要（英）：

The invention provides a double digital PCR fluorescent quantitative detection method for transgenic maize NK603. According to the method, quantitative detection can be directly performed on the transgenic maize NK603 in genomes of samples to be detected, and therefore the standard curve plotting step in quantitative detection of a conventional PCR method and the sample preprocessing step in existing digital PCR detection are omitted; in addition, transgenic ingredients are calculated according to the proportion of the exogenous gene copy number and the reference gene copy number, and therefore the stability of quantitative detection of the transgenic ingredients can be improved by performing double detection on one reaction system. Through the method, absolute quantitative detection on the transgenic maize NK603 can be achieved, the quantitative detection limitation can reach 0.5%, the sensitivity can reach 0.1%, and the requirements of actual detection of all the transgenic ingredients can be met. In addition, the method is easy to operate and high in flexibility and serves as an effective method for absolute quantitative detection of transgenes.

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