| 韩蕊，滕祎，李献锋，王临静，魏霜，张雍哲，乔曦，刘海军，吴尧，马骏，万方浩，刘聪辉，钱万强,2024,基于酶介导双重指数扩增技术（EmDEA）的小火蚁快速检测方法的开发[J].环境昆虫学报,(5):1059-1067 |
| 基于酶介导双重指数扩增技术（EmDEA）的小火蚁快速检测方法的开发 |
| Development of a rapid detection method for Wasmannia auropunctata based on enzyme-mediated duplex exponential amplification (EmDEA) |
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| DOI： |
| 中文关键词: 小火蚁 酶介导双重指数扩增技术（EmDEA） 特异性引物 RNA探针 快速鉴定 精准检测 |
| 英文关键词:Wasmannia auropunctata EmDEA specific primers RNA probes rapid identification precise detection |
| 基金项目:国家重点研发计划（2022YFC2601500） |
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| 中文摘要: |
| 小火蚁Wasmannia auropunctata是危害性最严重的入侵蚂蚁之一，对动植物、生态、经济甚至是人类都容易造成严重影响，是我国的重大检疫害虫之一。本研究为实现小火蚁的现场快速精准检测，通过自研程序筛选出小火蚁全基因组中的特异性片段，具有更高的特异性，基于酶介导双重指数扩增技术（EmDEA）筛选了适用的引物、RNA探针，全部反应所需时间小于20 min，并验证了其特异性和灵敏度。结果显示这种方法可在短时间内达到极高检测灵敏度与特异性，最低检出限约为4 ng。这种方法极大程度降低对设备的依赖，简单、快速、特异，为小火蚁的基层检测与现场诊断提供技术支持。 |
| 英文摘要: |
| Wasmannia auropunctata is one of the most harmful invasive ants, posing severe threats to flora, fauna, ecosystems, economies, and even humans. It is a significant quarantine pest in China. To achieve rapid and precise on-site detection of the W. auropunctata, this study screened specific segments from the whole genome of the W. auropunctata through self-developed program, ensuring higher specificity. The study also selected suitable primers and RNA probes based on the enzyme-mediated duplex exponential amplification (EmDEA). The total reaction time required was less than 30 minutes, and both specificity and sensitivity were validated. Results demonstrate that this method could achieve extremely high detection sensitivity and specificity in a short time, with a minimum detection limit of approximately 4 ng. This method significantly reduces the dependence on equipment, making it simple, rapid, and specific, thereby providing technical support for grassroots detection and on-site diagnosis of the W. auropunctata. |
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