An enhanced qPCR method for rapid Agrilus planipennis detection and monitoring

Emerald ash borer (EAB; Agrilus planipennis) represents a serious threat to North American and European ash species (Fraxinus spp.). Spread of EAB westwards, from European Russia and Eastern Ukraine, could lead to dramatic consequences for native European ash populations. Early detection is essential for fast and successful eradication of new populations. In this study, we developed a new TaqMan qPCR assay allowing for sensitive and specific detection of EAB. We tested the specificity of the assay against 17 European Agrilus spp., eight buprestid species and nine species belonging to other wood-associated beetle taxa. The qPCR assay provided reliable amplification from samples with DNA concentrations as low as 0.5 picograms per reaction. Moreover, DNA could be amplified from different sample types, such as egg casings, leaves, faeces and bore dust from larval galleries. Robustness of the assay was verified by performing a blind test with four different laboratories. Here we provide a highly specific, robust and sensitive assay which can be used for enhanced surveillance of Agrilus planipennis on the European continent.