Research project Fast and harmonised detection methods for quarantine Tephritidae

In progress TEPHRIFADE 2 01/06/2026 → 31/05/2028

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Negin Ebrahimi

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General introduction

Non-EU Tephritidae fruit flies are among the most damaging pests of fruit and vegetable crops and represent a major threat to EU agriculture and trade. More than 250 non EU Tephritidae species are regulated in the EU, including priority pests such as Anastrepha ludens and Bactrocera dorsalis. Rapid and reliable identification is essential for quarantine inspections, but current molecular tools are available for only a limited number of species. In a previous project (TEPHRIFADE 1) important gaps in rapid diagnostics, harmonised protocols and DNA reference databases were revealed. However, rapid and reliable identification is essential for inspections of these quarantine organisms, while current molecular tools are available only for a limited number of species, which prevents laboratories from identifying intercepted specimens quickly and consistently. More than 250 non-EU Tephritidae species are regulated in the EU, including priority pests such as Anastrepha ludens and Bactrocera dorsalis. This project will improve preparedness, surveillance, and phytosanitary decision-making for high-risk fruit fly pests.

Research approach

Specifically, within TEPHRIFADE 2 rapid LAMP or qPCR assays for Anastrepha ludens and A. fraterculus will be developed and validated, supported by genomic data, including COI barcodes, mitochondrial genomes and nuclear SNP markers. The researchers optimise an existing TaqMan real-time PCR assay for Bactrocera dorsalis and explore olfactory gene-based markers for the B. dorsalis species complex. MALDI TOF MS will be assessed as a complementary tool for larval identification of A. ludens and A. fraterculus. Rapid DNA extraction methods that preserve morphology will be evaluated. A harmonised validation framework, workshop and Test Performance Studies will support interlaboratory comparison. DNA reference gaps will be addressed through data mining, museum and institutional collections, DNA repositories and targeted field sampling.

Relevance/Valorization

TEPHRIFADE 2 is expected to strengthen European diagnostic capacity for regulated non-EU Tephritidae by delivering faster, more reliable and harmonised identification tools. The project will support national plant protection organisations and diagnostic laboratories with rapid responses to to interceptions and suspected outbreaks. Validated assays, standardised extraction protocols and Test Performance Studies will improve consistency between laboratories. Expanded DNA reference data will increase the reliability of molecular identification and support future diagnostic development. Collaboration with EURL and EPPO will facilitate alignment with European diagnostic standards.