Strengthening hazelnut certification systems through sensitive detection of apple mosaic virus (ApMV)

Abstract Utilizing virus-free materials is one of the most effective strategies for managing plant viruses. Establishing a robust phytosanitary certification program is the primary method to inhibit yield losses caused by the apple mosaic virus (ApMV) in hazelnuts by providing growers with selected propagation materials. Currently, the hazelnut certification system 2000/29/EC relies solely on Double Antibody Sandwich Enzyme-Linked Immunosorbent Assay (DAS-ELISA) and conventional Reverse Transcription Polymerase Chain Reaction (RT-PCR) for diagnosing ApMV. Accurate and prompt detection of ApMV infection in host plants is crucial for effective disease management. This paper presents a new study aimed to assess the sensitivity of various diagnostic tests for detecting ApMV in rootstock hazelnut plants (Kara and Foşa), comparing DAS-ELISA, RT-PCR, and real-time RT-PCR (RT-qPCR). The findings indicate that RT-qPCR is the most effective and practical method for screening ApMV in hazelnuts and apples, as it can overcome false negatives associated with both ELISA tests and RT-PCR amplification. While DAS-ELISA showed limited sensitivity in basic hazelnut seedlings, RT-PCR achieved a detection limit of 100 pg/μL, and RT-qPCR reached a more sensitive detection limit of 100 fg/μL. The results of this study are expected to enhance the application of existing methods in ApMV diagnosis and support virus-free hazelnut certification.