TEPHRIFADE 2 builds on the previous TEPHRIFADE 1 project to strengthen diagnostic capacity for quarantine Tephritidae. The project will develop rapid molecular identification methods, such as LAMP or qPCR, for Anastrepha fraterculus and A. ludens, and will evaluate MALDI TOF mass spectrometry as a complementary identification approach. It will also establish a framework for validating and harmonising molecular diagnostic methods across laboratories, including updated diagnostic method lists, a diagnostic workshop, and test performance studies. In parallel, the project will address gaps in reference data by targeting non EU Tephritidae species for which COI barcodes are still missing, using specimen collection, museum material, Sanger sequencing and whole genome sequencing where appropriate. RMCA contributes Tephritidae taxonomic expertise, genomic resources, molecular marker discovery and support for the development and validation of diagnostic tools.