Ultrasensitive, low-input detection of avocado sunblotch viroid via RPA-CRISPR and nanopore-array single-bead fluorescence readout.
Xu J, Jiang X, Dashtarzhaneh MK, Zhong Y, Sharma B
Crispr
Avocado trees infected with Sunblotch Viroid can look perfectly healthy for years while quietly producing stunted, mottled fruit and spreading the pathogen to every tree grown from their cuttings — with no cure once it takes hold.
Avocado Sunblotch Viroid is a tiny, invisible pathogen that causes avocado trees to produce deformed, discolored fruit and can silently spread through an orchard for years before anyone notices. Researchers built a new test combining two powerful molecular tools — one that copies trace amounts of the viroid's genetic material and another that acts like molecular scissors to signal its presence — to detect even the tiniest infection from a small piece of leaf or bark. The whole test takes just one hour, needs no big lab machines, and could eventually be used right in an orchard to catch the disease early enough to contain it.
Key Findings
Detected Avocado Sunblotch Viroid at concentrations as low as 1.68 copies per microliter, representing one of the most sensitive viroid detection thresholds reported.
Used only 40 nanoliters of sample per readout — more than 100-fold less plant material than conventional fluorescence-based assays require.
The complete test runs in 60 minutes at a constant temperature with no PCR thermocycler needed, enabling potential deployment directly in orchards or the field.
chevron_right Technical Summary
Scientists developed an ultra-sensitive, portable test for Avocado Sunblotch Viroid that can detect the pathogen at fewer than 2 copies per microliter from a tiny drop of plant extract — making early orchard diagnosis practical without a laboratory.
Abstract Preview
Rapid and sensitive detection of plant pathogens, such as the Avocado Sunblotch Viroid (ASBVd), is essential for early disease management and agricultural biosecurity. Yet, most current diagnostic ...
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