刘二龙,卢丽,吕英姿,蒋湘,李立霞,李嘉琪,杜雅萍,郑高彬.转基因鲑鱼AquAdvantage品系特异性实时荧光聚合酶链式反应检测方法的建立[J].中国食品卫生杂志,2017,29(5):576-580. 本文二维码信息
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转基因鲑鱼AquAdvantage品系特异性实时荧光聚合酶链式反应检测方法的建立
Event-specific real-time polymerase chain reaction detection method of genetically modified AquAdvantage salmon
投稿时间:2017-05-05  
DOI:
中文关键词:  转基因鲑鱼  AquAdvantage  实时荧光聚合酶链式反应  品系特异性  检测
Key Words:Transgenic salmon  AquAdvantage  real-time polymerase chain reaction  event-specific  detection
基金项目:广东出入境检验检疫局科技项目(2017GDK41)
作者单位E-mail
刘二龙 黄埔出入境检验检疫局,广东 广州 510730 erlongliu@126.com 
卢丽 广东出入境检验检疫局技术中心,广东 广州 510623  
吕英姿 黄埔出入境检验检疫局,广东 广州 510730  
蒋湘 黄埔出入境检验检疫局,广东 广州 510730  
李立霞 黄埔出入境检验检疫局,广东 广州 510730  
李嘉琪 黄埔出入境检验检疫局,广东 广州 510730  
杜雅萍 黄埔出入境检验检疫局,广东 广州 510730  
郑高彬 黄埔出入境检验检疫局,广东 广州 510730  
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中文摘要:
      目的 实现转基因鲑鱼AquAdvantage的标识管理,建立其品系特异性实时荧光聚合酶链式反应(PCR)检测方法。方法 针对转基因鲑鱼的品系特异性序列设计引物和TaqMan探针,建立转基因鲑鱼实时荧光PCR检测方法,并对该方法的特异性、灵敏度和重复性进行检测。结果 建立的转基因鲑鱼实时荧光PCR方法特异性强,在600 000~60拷贝范围内呈良好的线性关系,其线性回归方程为y=-3.2194x+40.805,R2=0.997,检测限为60拷贝,检测重复性良好。结论 建立的品系特异性实时荧光PCR方法可应用于转基因鲑鱼AquAdvantage的鉴定。
Abstract:
      Objective For implementation of labeling regulations, an event-specific real-time polymerase chain reaction(PCR) method for the detection of genetically modified AquAdvantage salmon was established in this study. Methods Primers and TaqMan probe were designed based on the event-specific sequence of AquAdvantage salmon. The specificity, sensitivity and repeatability of the developed method were examined, respectively. Results The specificity test of this method showed it was specific to AquAdvantage salmon. The 600 000-60 copies range showed a good linear relationship with Ct values, and its linear regression equation was y=-3.2194x+40.805(R2=0.997). The limit of quantification(LOQ) was 60 copies and the repeatability was good. Conclusion This event-specific real-time PCR method was suitable for the identification of genetically modified AquAdvantage salmon.
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