摘要
目的为实现转基因甜菜GTSB77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GTSB77的3’端外源插入片段与甜菜基因组DNA之间的邻接区序列设计引物和探针,建立GTSB77品系特异性实时荧光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GTSB77检测方法特异性强,定量限(LOQ)为16拷贝,扩增效率为102%,重复性测试结果相对标准偏差(RSD)介于0.21%~1.66%之间。结论建立的实时荧光PCR方法可应用于GTSB77的鉴定检测。
Objective For implementation of labeling regulations,an event-specific real-time polymerase chain reaction(PCR)method for quantitative detection of sugarbeet GTSB77 was established.Methods The specific primer pairs and probe based on the 3’flanking sequence of GTSB77 were designed.The specificity,sensitivity and repeatability of the developed method were examined,respectively.Results The specificity test showed it specific for GTSB77 detection.The limit of quantification(LOQ)was 16 copies and the amplification efficiency was 102%.Reapeatability of the established method was assessed and the relative standard deviation(RSD)ranged from 0.21%-1.66%.Conclusion This eventspecific real-time PCR method is suitable for identification of GTSB77.
作者
刘二龙
卢丽
吕英姿
蒋湘
李嘉琪
夏柔菲
LIU Erlong;LU Li;LYU Yingzi;JIANG Xiang;LI Jiaqi;XIA Roufei(Huangpu Customs Technology Center,Guangdong Guangzhou 510730,China;Guangzhou Customs Technology Center,Guangdong Guangzhou 510623,China)
出处
《中国食品卫生杂志》
CSCD
北大核心
2020年第1期49-52,共4页
Chinese Journal of Food Hygiene
基金
广东出入境检验检疫局科技项目(2018GDK50).
