ArticlePlant biotechnology journal2021
Field detection of multiple RNA viruses/viroids in apple using a CRISPR/Cas12a-based visual assay.
Article in Plant biotechnology journal, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 58 papers.
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Who cites it
58 citing papers in PubMed, 156 citations in OpenAlex.
- Attomolar Detection of HIV-1 With Label-Free RCA-rCRISPR on Smartphone.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Review
- Field-parallel six-sample microfluidic detection of plant viruses via raffinose-assisted one-pot LAMP-CRISPR/Cas12b.Journal of advanced research · 2026Article
- CRISPR-Cas systems for plant virus management: detection, surveillance, and host resistance.Frontiers in plant science · 2026Review
- Emerging technologies for in-field plant virus detection: innovations and future directions.The Journal of general virology · 2025Review
- Managing Plant Viruses in Tissue-Cultured Apple and Grapevine: Strategies for Detection and Eradication.The plant pathology journal · 2025Review
- Transmission of apple stem grooving virus (Capillovirus mali) to apple from the soil-borne fungus Fusarium solani.BMC plant biology · 2025Article
- Detection of tomato brown rugose fruit virus through CRISPR-Cas12a and CRISPR-Cas9 systems.Scientific reports · 2025Article
- CRISPR-Cas12a/RPA Dual-Readout Assay for Rapid Field Detection of Porcine Rotavirus with Visualization.Viruses · 2025Article
- Detection of TuMV by a toehold switch sensor coupled with NASBA amplification in Pseudostellaria heterophylla.Plant methods · 2025Article
- Conventional and cutting-edge advances in plant virus detection: emerging trends and techniques.3 Biotech · 2025Review
- Single and dual RPA-CRISPR/Cas assays for point-of-need detection of Stewart's wilt pathogen (Pantoea stewartii subsp. stewartii) of corn and Maize dwarf mosaic virus.Pest management science · 2025Article
- CRISPR-Cas-Based Diagnosis of Geminiviruses.Methods in molecular biology (Clifton, N.J.) · 2025Article
- RPA-assisted CRISPR-Cas12a-enabled point-of-care diagnostic platform for chili leaf curl virus with fluorescent and colorimetric readouts.Frontiers in microbiology · 2025Article
- Unveiling viral threats to temperate pome fruits: characterization, transmission, and sustainable management strategies.Frontiers in microbiology · 2025Review
- CRISPR-Cas-Based Pen-Side Diagnostic Tests forMicroorganisms · 2024Article
- Article
- Applications of CRISPR Technologies in Forestry and Molecular Wood Biotechnology.International journal of molecular sciences · 2024Review
- A novel system with robust compatibility and stability for detectingMicrobiology spectrum · 2024Article
- Advances in plant pathogen detection: integrating recombinase polymerase amplification with CRISPR/Cas systems.3 Biotech · 2024Review
Corrections and comments
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Authors and funding
12 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Co-infection of apple trees with several viruses/viroids is common and decreases fruit yield and quality. Accurate and rapid detection of these viral pathogens helps to reduce losses and prevent virus spread. Current molecular detection assays used for apple viruses require specialized and expensive equipment. Here, we optimized a CRISPR/Cas12a-based nucleic acid detection platform for the diagnosis of the most prevalent RNA viruses/viroid in apple, namely Apple necrotic mosaic virus (ApNMV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus (ACLSV) and Apple scar skin viroid (ASSVd). We detected each RNA virus/viroid directly from crude leaf extracts after simultaneous multiplex reverse transcription-recombinase polymerase amplification (RT-RPA) with high specificity. Positive results can be distinguished by the naked eye via oligonucleotide-conjugated gold nanoparticles. The CRISPR/Cas12a-RT-RPA platform exhibited comparable sensitivity to RT-qPCR, with limits of detection reaching 250 viral copies per reaction for ASPV and ASGV and 2500 copies for the others. However, this protocol was faster and simpler, requiring an hour or less from leaf harvest. Field tests showed 100% agreement with RT-PCR detection for 52 samples. This novel Cas12a-based method is ideal for rapid and reliable detection of apple viruses in the orchard without the need to send samples to a specialized laboratory.
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Registered trials
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