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基于短DNA适配体的灵敏比色适配体传感器的研制及其在农业土壤中百草枯残留筛查中的应用。

Development of sensitive colorimetric aptasensor based on short DNA aptamer and its application to screening for paraquat residues in agricultural soil.

作者信息

Kongpreecha Pakawat, Phanchai Witthawat, Puangmali Theerapong, Siri Sineenat

机构信息

School of Biology, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima, Thailand.

Department of Physics, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand.

出版信息

Biotechnol Appl Biochem. 2023 Apr;70(2):857-869. doi: 10.1002/bab.2405. Epub 2022 Sep 27.

DOI:10.1002/bab.2405
PMID:36112641
Abstract

Paraquat is a widely used herbicide for controlling weeds and grasses in agriculture, and its contaminated residues in agricultural areas are of increasing concern. This work reports the development of the sensitive and easy-to-use colorimetric aptasensor for screening paraquat residues in agricultural soil. The short DNA fragments derived from the original aptamer were analyzed for their capability to interact with paraquat by molecular dynamic simulation. The paraquat-aptasensor was developed using the selected DNA fragment and gold nanoparticles. Its limit of detection (LOD) for paraquat is 2.76 nM, which is more sensitive than the aptasensor with long-length aptamer (LOD = 12.98 nM). The developed aptasensor shows the selectivity to paraquat, but not to other tested herbicides; ametryn, atrazine, difenzoquat, 2,4-D-dimethyl ammonium, and glufosinate. The recovery rates of paraquat detection in the spiked soil samples were in a range of 99.5%-105.1%, with relative standard deviation values of <4%. The developed aptasensor was used to screen for paraquat residues in agricultural soils, and three out of 23 soil samples were tested positive for paraquat, which was confirmed by a high-performance liquid chromatography analysis. These results suggested the potential application of the developed aptasensor to detect paraquat residues in agricultural sites.

摘要

百草枯是一种广泛用于农业中控制杂草的除草剂,其在农业区域的污染残留日益受到关注。本研究报告了一种灵敏且易于使用的比色适体传感器的开发,用于筛选农业土壤中的百草枯残留。通过分子动力学模拟分析了源自原始适体的短DNA片段与百草枯相互作用的能力。利用所选DNA片段和金纳米粒子开发了百草枯适体传感器。其对百草枯的检测限(LOD)为2.76 nM,比具有长链适体的适体传感器(LOD = 12.98 nM)更灵敏。所开发的适体传感器对百草枯具有选择性,但对其他测试除草剂(如莠灭净、阿特拉津、敌草快、2,4-D-二甲基铵和草铵膦)没有选择性。加标土壤样品中百草枯检测的回收率在99.5%-105.1%范围内,相对标准偏差值<4%。所开发的适体传感器用于筛选农业土壤中的百草枯残留,23个土壤样品中有3个经高效液相色谱分析确认对百草枯检测呈阳性。这些结果表明所开发的适体传感器在检测农业场地百草枯残留方面具有潜在应用价值。

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