多物种复合样品中转基因DNA成分的定量分析  

Quantification of DNA from genetically modified materials by real-time PCR

作　　者：杨冬燕[1] 侯红利[2] 杨永存[1] 周向阳[2] 杨小柯[1] 郭良让[3] 李永红[2] 林健荣[3] 邓平建[1] 

机构地区：[1]广东省深圳市疾病预防控制中心,广东深圳518020 [2]广东省深圳市农作物良种引进中心,广东深圳518040 [3]华南农业大学,广州510642

出　　处：《中国卫生检验杂志》2009年第8期1727-1730,共4页Chinese Journal of Health Laboratory Technology

基　　金：广东省自然科学基金资助项目(06027682);深圳市科技研发资金([2007]334-9)

摘　　要：目的：在前期研究建立的不依赖参比内源基因定量单物种植物样品中转基因DNA的方法的基础上,定量测定多物种复合样品中转基因DNA的含量。方法：以转基因和非转基因大豆、玉米、大米按比例混合配制的多物种复合材料为研究对象,通过实时荧光PCR法测定样品DNA提取物中转基因DNA量,通过测定260 nm处的吸光值测定样品DNA提取物中总DNA量,以样品DNA提取物中各物种转基因DNA量与总DNA量之比表示多物种复合样品中各物种转基因DNA成分含量,并以各物种转基因DNA成分含量之和计算样品中转基因DNA的总含量。结果：7个大米与玉米两物种混合样品和7个大米、大豆与玉米三物种混合样品用于验证新建的定量体系的准确度和精密度。结果表明,两物种和三物种复合样品转基因成分总含量测定值与理论值的偏离度分别为2.22%-19.54%和5.24%-20.51%;其转基因成分总含量测定的变异系数分别为2.11%-13.43%和0.56%-10.74%。结论：新建的多物种复合样品中转基因DNA的含量测定方法满足转基因成分定量分析准确度和精密度要求。Objective： To establish a method to quantify genetically modified（GM） content in composite GM materials and to validate its precision and closeness.Methods：On one hand real-time PCR to determine the amount of GM targets present and on the other hand absorbance reading at 260nm to measure the total DNA present in sample was used in the study.Results：The quantitative results derived from a total of 14 GM DNA composite samples containing defined proportions of GM materials.The bias for the quantification of the total GM content in two-species and three-species composite samples ranged from 2.22% to 19.54% and from 5.24% to 20.51% respectively.For 7 of two-species composite samples and 7 of three-species composite samples,the coefficients of variation（CV） were respectively from 2.11% to 13.43% and from 0.56% to 10.74%.Conclusion：The quantitative method is a feasible assay to quantify GM DNA in composite materials in term of precision and closeness to the expected value.

关 键 词：转基因 定量 复合样品 实时荧光PCR 

分 类 号：R319[医药卫生—基础医学]