Development of a novel LAMP assay for early detection of two-line chestnut borer (Agrilus bilineatus)

Sammanfattning

The two-line chestnut borer (Agrilus bilineatus) poses a major threat to oak populations in Europe. It is a close relative to the invasive emerald ash borer (Agrilus planipennis), known for its catastrophic impact on ash (Fraxinus spp.) forest. With increase in global trade, the threat of A.bilineatus spreading further into Europe is a serious concern for forest health and biosecurity. Quick and reliable molecular detection methods hence are needed to identify these pests before they get established. In this study, I aim to create a loop isothermal amplification assay for the early detection of TLCB. I have tested the assay against 24 non-target species, comprising of 16 Agrilus species (Buperstidae), three non-Agrilus buprestids, four Cerambycids, and two Curculionids. The assay was specific to the target species showing clear amplification and no amplification for all non-target species. The assay was also tested for sensitivity, and it successfully amplified DNA as low as to 0.064 pg/μL. The result demonstrates high specificity and sensitivity of the assay providing a reliable tool for early detection and can be used on-site to monitor the presence of forest pests enabling proactive management of forest pest invasions.