ARPHA Preprints, doi: 10.3897/arphapreprints.e163096

An enhanced qPCR method for rapid Agrilus planipennis detection and monitoring

Quirin Kupper^‡, Donnie Lee Peterson^§, Leandra Carina Fritsi^|, Doris Hölling^¶, Anouchka Perret-Gentil^‡, Francesco Pecori^#, Denise Altenbach^¤, Marco Giulio^«, Helen Zbinden^», Salome Schneider^|, Beat Ruffner^‡

‡ Swiss Federal Research Institute for Forest, Snow and Landscape, WSL, Birmensdorf, Switzerland§ Swedish University of Agricultural Sciences, Alnarp, Sweden| Forest Health and Biotic Interactions, Swiss Federal Research Institute for Forest, Snow and Landscape, Birmensdorf, Switzerland¶ WSL, Birmensdorf, Switzerland# CNR, Sesto fiorentino (FI), Italy¤ Diagnostic moléculaire des organismes nuisibles réglementés des végétaux, Agroscope, Nyon, Switzerland« 5 Department of Aquatic Ecology, Swiss Federal Institute of Aquatic Science and Technology (Eawag), Dübendorf, Switzerland» Department of Plant and Microbial Biology, University of Zurich, Zurich, Switzerland

Corresponding author: Quirin Kupper ( quirin.kupper@wsl.ch )

Corresponding author: Beat Ruffner ( beat.ruffner@wsl.ch )

This is an open access preprint distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Citation: Kupper Q, Peterson DL, Fritsi LC, Hölling D, Perret-Gentil A, Pecori F, Altenbach D, Giulio M, Zbinden H, Schneider S, Ruffner B (2025) An enhanced qPCR method for rapid Agrilus planipennis detection and monitoring. ARPHA Preprints. https://doi.org/10.3897/arphapreprints.e163096

Abstract

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 five picograms per reaction. Moreover, DNA could be amplified from different sample types such as egg casings, leaves, feces 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.

Keywords

Agrilus planipennis, Emerald Ash Borer, EAB, invasive forest insects, TaqMan qPCR, surveillance