一个基于LAMP-CRISPR-Cas12a的快速和高通量检测转基因产品的系统
Hua Liu1, Xiuwen Hu2, Haijuan Zeng1
1Institute of Biotechnology Research, Shanghai Academy of Agricultural Sciences, Key Laboratory of Agricultural Genetics and Breeding, 2901 Beidi Road, Shanghai, 201106, China.
Current research in food science
|October 23, 2023
概括
一种新的循环介导同热放大 (LAMP),CRISPR-Cas12a和侧向流量检测 (LFA) 方法可以在田间快速,灵敏地检测转基因作物基因. 这项技术可用于监管目的快速选转基因生物.
科学领域:
- 农业生物技术 农业生物技术
- 分子生物学分子生物学
- 生物传感技术的技术
背景情况:
- 全球转基因作物的增加需要有效的监管和选检测方法.
- 现有的转基因检测技术可能缺乏现场应用所需的速度,灵敏度或可移植性.
研究的目的:
- 开发一种快速,灵敏和特定的GM作物基因现场检测方法.
- 建立一个用户友好的系统,用于对转基因生物 (GMO) 的现场查.
主要方法:
- 开发一个结合循环介导同热放大 (LAMP),CRISPR-Cas12a和侧向流量测试 (LFA) 系统.
- 反应条件的优化和原料选,以提高特异性和灵敏度.
- 使用含有特定转基因基因 (CP4-EPSPS和Cry1Ab/Ac) 的实地测试样本验证该方法.
主要成果:
- 通过LAMP-CRISPR-Cas12a-LFA方法,使用横流测试条实现了100个副本的检测极限 (LOD).
- 在61°C的恒温下,整个检测过程在1小时内完成.
- 该系统表现出高可重现性,并在现场样本中成功检测出目标基因.
结论:
- 开发的LAMP-CRISPR-Cas12a-LFA方法为在现场检测转基因作物基因提供了快速可靠的工具.
- 它的便携性和易用性使其适用于各种应用,包括实验工作站,执法部门和检查部门.
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