转基因作物多重重组酶聚合酶扩增结合CRISPR/Cas12a现场可视化检测方法的建立(英文)(Establishment of a field visualization detection method for multiplex recombinase polymerase amplification combined with CRISPR/Cas12a in genetically modified crops)

转基因作物多重重组酶聚合酶扩增结合CRISPR/Cas12a现场可视化检测方法的建立(英文)(Establishment of a field visualization detection method for multiplex recombinase polymerase amplification combined with CRISPR/Cas12a in genetically modified crops)

. With the approval of more and more genetically modified (GM) crops in our country, GM safety management has become more important. Transgenic detection is a major approach for transgenic safety management. Nevertheless, a convenient and visual technique with low equipment requirements and high sen...

Full description

Saved in:
Bibliographic Details
Main Authors: 颜晶莹(YAN Jingying), 倪亮(NI Liang), 沈星宇(SHEN Xingyu), 吕秉韬(LÜ Bingtao), 李玉(LI Yu)
Format: Article
Language:English
Published: Zhejiang University Press 2025-06-01
Series:浙江大学学报. 农业与生命科学版
Subjects:
转基因作物
重组酶聚合酶扩增
crispr/cas12a
现场检测
Online Access:https://doi.org/10.3785/j.issn.1008-9209.2024.01.162
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:. With the approval of more and more genetically modified (GM) crops in our country, GM safety management has become more important. Transgenic detection is a major approach for transgenic safety management. Nevertheless, a convenient and visual technique with low equipment requirements and high sensitivity for the field detection of GM plants is still lacking. On the basis of the existing recombinase polymerase amplification (RPA) technique, we developed a multiplex RPA (multi-RPA) method that can simultaneously detect three transgenic elements, including the cauliflower mosaic virus 35S gene (CaMV35S) promoter, neomycin phosphotransferase Ⅱ gene (NptⅡ) and hygromycin B phosphotransferase gene (Hyg), thus improving the detection rate. Moreover, we coupled this multi-RPA technique with the CRISPR/Cas12a reporter system, which enabled the detection results to be clearly observed by naked eyes under ultraviolet (UV) light (254 nm; which could be achieved by a portable UV flashlight), therefore establishing a multi-RPA visual detection technique. Compared with the traditional test strip detection method, this multi-RPA-CRISPR/Cas12a technique has the higher specificity, higher sensitivity, wider application range and lower cost. Compared with other polymerase chain reaction (PCR) techniques, it also has the advantages of low equipment requirements and visualization, making it a potentially feasible method for the field detection of GM plants.
ISSN:2097-5155

Similar Items