Novel Agrilus planipennis (Coleoptera: Buprestidae) early-detection tool designed on cytochrome B gene

Agrilus planipennis (Coleoptera: Buprestidae) is a quarantine pest that is threatening native ash populations in both North America and Europe. Early detection of this pest, which can cause severe infestations, is essential, and molecular tests applied at different developmental stages and environmental traces can contribute substantially to improving control measures to prevent or contain A. planipennis infestations. In this study, a specific A. planipennis real-time qPCR assay was developed using a Locked Nucleic Acid (LNA) probe based on the CytB (cytochrome B) gene. The detection limit of this method was 25.6 fg/µl for adult DNA extracts and 0.21 pg/µl for frass produced by A. planipennis larvae. The new qPCR probe test, which targets a different locus, not only allows identification of the pest and provides an indirect diagnosis through environmental DNA analysis but can also be used for cross-validation of results between different tests.