植物DNA混合物中物种成分实时荧光PCR定量分析  

Plant species quantification in DNA mixtures by real-time polymerase Chain reaction

作　　者：杨永存[1] 杨冬燕[1] 王学林[2] 杨小柯[1] 周向阳[2] 郭良让[3] 林健荣[3] 邓平建[1] 吴水清[2] 

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]华南农业大学动物科学学院,广州510642

出　　处：《中国卫生检验杂志》2009年第6期1206-1209,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);2007年深圳市科技研发基金(深科信[2007]334号-9)

摘　　要：目的：建立植物DNA混合物中物种成分实时荧光PCR定量方法。方法：建立PCR反应管中某物种DNA质量和该物种特异性基因扩增Ct值之间的定量关系,通过260 nm处光密度值的测定计算DNA提取液的浓度和DNA总质量,并以某物种DNA和总DNA的质量百分比表示该物种DNA含量。对56个待测样品中的物种成分进行定量测定以验证新建方法的准确性和精密度。结果：新建方法的LOQ低于0.51%;B ias为0.00%～20.79%,RSD为0.43%～19.64%。结论：新建方法的准确度和精密度均符合现行要求。Objective： To develop a model for plant species quantification in DNA mixtures by real - time polymerase chain reaction. Methods： Direct quantitative relation between the DNA mass of a certain species in PCR reaction and the Ct values of corresponding species - specific gene was established. By the measurement of optical density at 260 -nm ultraviolet ray （UV） , DNA concentration and the total DNA mass of DNA extract was obtained. The content of a certain species DNA was expressed as the mass% of target species DNA to the total DNA in sample. The content of a certain species DNA in 56 tested samples were quantified to estimate the accuracy and precision of the novel model. Results ： The limit of quantification （LOQ） of the novel model was lower than 0. 51%. The bias varied from 0.00% to 20. 79%, and the repeatability standard deviation （RSD） varied from 0. 43% to 19.64%. Conclusion： Both the accuracy and precision of the novel model meet the exist requirements.

关 键 词：DNA混合物 物种定量 实时荧光PCR 

分 类 号：R446[医药卫生—诊断学]