Primary Industries Development, Department of Primary Industries and Regional Development, 3 Baron-Hay Court, Kensington, Western Australia, 6151, Australia.
Sustainability and Biosecurity, Department of Primary Industries and Regional Development, 3 Baron-Hay Court, Kensington, Western Australia, 6151, Australia.
J Econ Entomol. 2021 Dec 6;114(6):2610-2614. doi: 10.1093/jee/toab168.
Fall armyworm, Spodoptera frugiperda J.E. Smith (Lepidoptera: Noctuidae), is an economically important pest worldwide and has recently been identified in Australia. Morphological identification of S. frugiperda at early larval stages can be difficult often requiring expert microscopy analysis. Rapid and accurate in-field diagnosis is vital for management decision support and there are no tools currently available for this purpose. In this study, a sensitive, specific, and in-field capable loop-mediated isothermal amplification (LAMP) assay was developed to detect S. frugiperda larvae. A primer set based on a highly conserved region of the S. frugiperda cytochrome oxidase subunit 1 (COX1) gene provided detection within 30 min from both total DNA and crude extractions. The crude extraction technique of crushing 10 mg of S. frugiperda material in 50 µl ddH2O and further diluting the homogenate in ddH2O is rapid, simple, and does not require heat blocks, centrifuges, or special buffers increasing its utility as a field-based technique. The primer set detected as little as 24 pg of S. frugiperda DNA and did not cross-react with any other of the lepidopteran species tested that are easily confused with S. frugiperda in Australia. Therefore, this assay could be used in-field to correctly identify the presence of S. frugiperda and thereby greatly assist with timely management decisions.
草地贪夜蛾，Spodoptera frugiperda J.E. Smith（鳞翅目：夜蛾科），是一种在世界范围内具有重要经济意义的害虫，最近在澳大利亚被发现。在早期幼虫阶段，S. frugiperda 的形态鉴定可能很困难，通常需要专家进行显微镜分析。快速准确的现场诊断对于管理决策支持至关重要，但目前尚无为此目的而设计的工具。在这项研究中，开发了一种灵敏、特异且适用于现场的环介导等温扩增（LAMP）检测方法，用于检测 S. frugiperda 幼虫。一组基于 S. frugiperda 细胞色素氧化酶亚基 1（COX1）基因高度保守区域的引物可在 30 分钟内从总 DNA 和粗提物中检测到 S. frugiperda。将 10 mg S. frugiperda 材料在 50 µl ddH2O 中粉碎并进一步将匀浆在 ddH2O 中稀释的粗提物提取技术快速、简单，且不需要热块、离心机或特殊缓冲液，从而增加了其作为现场技术的实用性。该引物组可检测低至 24 pg 的 S. frugiperda DNA，且与在澳大利亚容易与 S. frugiperda 混淆的任何其他鳞翅目物种均无交叉反应。因此，该检测方法可用于现场准确鉴定 S. frugiperda 的存在，从而极大地帮助及时做出管理决策。
