食品科学

• 分析检测 • 上一篇    下一篇

米饭和米酒中Bt基因快速检测方法的建立

索 娜,刘 佳,刘 梅,王雪强,王於建,肖红梅*,周光宏   

  1. 国家肉品质量安全质量控制工程技术研究中心,南京农业大学食品科技学院,江苏 南京 210095
  • 出版日期:2013-06-25 发布日期:2013-06-17

Rapid Detection of Bt Gene in Cooked Rice and Rice Wine

SUO Na,LIU Jia,LIU Mei,WANG Xue-qiang,WANG Yu-jian,XIAO Hong-mei*,ZHOU Guang-hong   

  1. National Meat Quality and Safety Control Engineering Technology Research Center, College of Food Science and Techndogy, Nanjing Agricultural University, Nanjing 210095, China
  • Online:2013-06-25 Published:2013-06-17

摘要:

建立米饭和米酒中Bt基因环介导等温扩增技术(LAMP)检测方法。以抗虫Bt基因(GeneBank登录号Y09787)为研究对象,利用LAMP技术,针对Bt基因的6个区域设计4条特异性引物,并对镁离子浓度、Bst DNA聚合酶添加量、反应温度、反应时间等扩增条件及体系的特异性、检测灵敏度进行探索。结果表明:成功建立了米饭和米酒的LAMP定性检测方法。所建立的方法能够简便、快速、有效的检测米饭和米酒中的Bt基因,检测限为0.005%。该LAMP检测方法具有高度的特异性及灵敏度,不需要特殊仪器,结果可靠。

关键词: Bt基因, 环介导等温扩增, 检测, 米制品

Abstract:

We developed a loop-mediated isothermal amplification (LAMP) method for the detection of Bt gene in cooked
rice and rice wine. Bt gene (GeneBank accession number Y09787) was amplified by a set of four special primers that
were designed according to six regions of the target gene. We optimized the reaction conditions Mg2+ concentration, Bst
DNA polymerase concentration, temperature and time. We also explored the specificity and sensitivity of the reaction. The
results showed that the LAMP reaction system allowed easy and effective detection of Bt gene with a high specificity and
sensitivity. The limit of detection was 0.005%. The LAMP detection method is specific, sensitive and reliable without need
for special instruments.

Key words: Bt gene, loop-mediated isothermal amplification (LAMP), detection, rice products