Abstract The emerald ash borer (EAB), Agrilus planipennis (Fairmaire), is the most destructive invasive insect species of ash (Fraxinus spp.) in North America. An accurate method for early detection of this noxious insect pest is indispensable to providing adequate warning of A. planipennis infestation. A loop-mediated isothermal amplification (LAMP) assay (EAB-LAMP) was developed based on mitochondrial cytochrome c oxidase subunit I (COI) gene. The EAB-LAMP required only 30 min at 65°C to amplify A. planipennis DNA from specimens collected from geographically distinct locations.There was no cross-reactivity with other Agrilus and insect species.The developed EAB-LAMP differentially detected traces of A. planipennis genome (COI) within frass from various Fraxinus species. EAB-LAMP was also able to distinguish among A. planipennis DNA and other Agrilus species and nontarget insect species in trap captures. By detecting A. planipennis DNA in two additional trap captures (in situ), the EAB-LAMP was more sensitive and reliable than visual inspection. We tested the quantitative nature of the assay by evaluating pooled trap samples and demonstrated that the EAB-LAMP was capable of functioning optimally using a pool size of at least five individual trap samples. This potentially circumvents the need to perform large-scale individual analysis for processing trap samples. Considering its performance, specificity, sensitivity, and repeatability, the developed EAB-LAMP could be a valuable tool to support strategy and operation of large-scale surveillance for A. planipennis and could profitably be used in routine monitoring programs for effective management of A. planipennis. Graphical Abstract

中文翻译：

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开发循环介导等温扩增分析作为检测翡翠灰螟（鞘翅目：Buprestidae）入侵的预警工具

摘要 翡翠灰螟（EAB）Agrilus planipennis（Fairmaire）是北美最具破坏性的灰螟（Fraxinus spp.）入侵昆虫物种。早期检测这种有毒害虫的准确方法对于提供对 A. planipennis 侵染的充分警告是必不可少的。基于线粒体细胞色素 c 氧化酶亚基 I (COI) 基因开发了环介​​导的等温扩增 (LAMP) 测定 (EAB-LAMP)。EAB-LAMP 在 65°C 下仅需要 30 分钟即可从不同地理位置采集的样本中扩增 A. planipennis DNA。与其他 Agrilus 和昆虫物种没有交叉反应。开发的 EAB-LAMP 差异检测到 A. planipennis 的痕迹。来自各种水曲柳物种的 frass 中的 planipennis 基因组 (COI)。EAB-LAMP 也能够区分 A. 陷阱捕获中的 planipennis DNA 和其他 Agrilus 物种和非目标昆虫物种。通过在两个额外的陷阱捕获（原位）中检测 A. planipennis DNA，EAB-LAMP 比目视检查更灵敏、更可靠。我们通过评估汇集的陷阱样本来测试该测定的定量性质，并证明 EAB-LAMP 能够使用至少五个单独的陷阱样本的池大小以最佳方式运行。这可能避免了为处理陷阱样品而进行大规模单独分析的需要。考虑到其性能、特异性、敏感性和可重复性，开发的 EAB-LAMP 可能是支持 A. planipennis 大规模监测策略和操作的宝贵工具，并且可以有利地用于常规监测计划，以有效管理 A. 扁豆。图形概要