Rapid detection methods for quarantine Tephritidae

General introduction

Early detection and identification of non-European quarantine Tephritidae species can help prevent the unwanted introduction of these insects into our country. Mapping and evaluating the (relatively new) rapid molecular methods for identifying these pests in imported fruits and vegetables is one of the main results ILVO researchers realized within the Euphresco project TEPHRIFADE, with 19 partners from 14 countries. Fruit flies of the family Tephritidae are among the most harmful invasive pest species in fruits and vegetables. They cause significant crop losses and pose a threat to producers and traders worldwide. It is therefore important to be able to identify them in all their life stages down to genus or species level.

Research approach

Now an inventory of rapid molecular methods for the identification of Tephritidae species has been made, including LAMP, qPCR, species-specific (multiplex) PCR and DNA barcoding. This showed that the rapid techniques were available for only a limited number of Tephritidae species. The project partners exchanged protocols, organized interlaboratory testing for validation and implementation of the best diagnostic protocols, and established guidelines to harmonize detection. Diagnostic protocols were developed for the molecular identification of Ceratitis cosyra using the LAMP assay and for Bactrocera cucurbitae using a Real-Time PCR assay. To identify taxonomic gaps, the researchers also collected DNA sequences from GenBank and BOLD databases, identifying 69 and 80 species without sequence data in GenBank and BOLD, respectively.

Relevance/Valorisation

Knowledge exchange events, including a three-day training workshop on high-throughput sequencing (HTS) and LAMP, organized by NVWA in collaboration with ILVO and the RMCA, provided practical lessons for project partners and external participants. A Test Performance Study (TPS) conducted by ILVO, in which 10 international laboratories participated, validated the effectiveness of the developed LAMP test for C. cosyra. Aiming for more efficient interception of these pests and a faster, more accurate detection of an outbreak, efforts were made to collect representative specimens from species for which DNA sequence data are not yet available. Unfortunately, this was not possible. To fill these gaps, we need to organize targeted collection efforts in regions where the missing species are endemic.

Funding provider(s)
FOD Volksgezondheid, Veiligheid van de voedselketen en Leefmilieu