P-193 - LAMP assay for improved detection of Avocado Sunblotch Viroid

Avocado Sunblotch Viroid (ASBVd) is a severe disease affecting avocado trees, leading to significant losses in yield, fruit quality, and tree vitality. Traditional detection methods such as RT-qPCR and TaqMan assays, while highly sensitive, require expensive equipment, trained personnel, and extensive processing time. To address these limitations, we used Loop-Mediated Isothermal Amplification (LAMP) protocol for ASBVd detection, offering a rapid, cost-effective, and highly sensitive alternative.  The optimized LAMP assay detected 1 copy/µl of the viroid, which is 1,000 times more sensitive than previously reported for conventional RT-PCR (880 copies/µl). The method was validated using avocado leaf samples collected from known infected trees, and the specificity was confirmed through comparison with established RT-PCR, RT-qPCR and sequence data from the RT-PCR product. The LAMP assay's efficiency, simplicity, and affordability make it a promising tool for early ASBVd detection in resource-limited settings and field applications, improving viroid management.