Spatial double-layer hydrogels enabled visual detection of Cladobotryum mycophilum based on recombinase-aided amplification - CRISPR/Cas12a
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Summary
This summary is machine-generated.A new hydrogel method, RCCH, rapidly detects cobweb disease pathogen Cladobotryum mycophilum. This advanced diagnostic tool enhances mushroom industry safety and disease monitoring.
Area Of Science
- Biotechnology
- Mycology
- Agricultural Science
Background
- Cobweb disease, caused by Cladobotryum mycophilum, leads to substantial losses in mushroom cultivation.
- Accurate and timely detection methods are crucial for managing this significant agricultural pathogen.
Purpose Of The Study
- To develop a novel, rapid, and sensitive detection method for Cladobotryum mycophilum.
- To integrate recombinase-aided amplification (RAA) with CRISPR/Cas12a technology in a hydrogel matrix for pathogen detection.
Main Methods
- A hydrogel-based system (RCCH) was designed, combining RAA with CRISPR/Cas12a detection.
- The method utilizes a porous PEG hydrogel for enzyme trapping and spatial co-localization of target DNA and CRISPR/Cas12a.
- Fluorescent signals generated by Cas12a's trans-cleavage activity enable visual detection.
Main Results
- The RCCH method achieved a low limit of detection (1 fg/μL) for C. mycophilum DNA.
- Exceptional specificity was observed against related fungi (Trichoderma spp.) and the host mushroom (Lentinula edodes).
- The entire detection process was completed in under 40 minutes.
Conclusions
- RCCH offers a rapid, accurate, and practical solution for detecting cobweb disease in mushroom cultivation.
- This innovative approach has the potential to significantly improve disease monitoring and enhance safety in the mushroom industry.
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