Diagnostic protocols for the detection of Acheta domesticus densovirus (AdDV) in cricket frass

•Two new quantitative and highly sensitive qPCR assays to detect Acheta domesticus densovirus (AdDV).•A novel, non-destructive AdDV screening protocol based on cricket faeces.•Improved tools for AdDV screening in commercial cricket operations and monitoring AdDV escape to wild cricket populations. The European house cricket (Acheta domesticus) is a species of interest for the emerging insect-as-food industry. Acheta domesticus densovirus (AdDV) is a member of the Parvoviridae virus family which infects A. domesticus, causing widespread mortality and even extinction of local cricket populations. Despite the well-known detrimental effects of AdDV in commercial rearing of A. domesticus there are no optimized protocols to accurately and non-destructively detect and quantify the virus. This study establishes a new protocol for the detection of AdDV in faecal material from A.domesticus. The protocol includes methodological improvements, such as upgrading from conventional PCR to quantitative real-time PCR and is much more sensitive than previously published protocols. Moreover, this study shows that cricket faeces are a suitable, non-destructive sample substrate to infer reliably if a cricket population is infected with AdDV or not. Early detection of lethal or economic threats, such as disease-causing viruses, is an essential part of commercial cricket management as well as for monitoring the risk of spread to wild cricket populations or to (human) consumers.