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Updated: Jul 12, 2025
DNA Virus Detection System Based on RPA-CRISPR/Cas12a-SPM and Deep Learning
Published on: May 10, 2024
Field-friendly and ultra-fast detection platform without nucleic acid extraction for virus detection
Gaihua Cao1, Yifan Xiong1, Yue Qiu1
1Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, 400044, PR China; State Key Laboratory of Cattle Diseases Detection (Chongqing) of Customs, Diagnosis and Testing Laboratory of Lumpy Skin Disease, Chongqing Customs Technology Center, Chongqing, 400020, PR China.
This study introduces an ultra-fast, nucleic acid-free detection platform for Lumpy Skin Disease Virus (LSDV). The novel system significantly reduces testing time and cost, offering a sensitive and portable solution for field diagnostics.
Area of Science:
- Molecular Biology
- Virology
- Biotechnology
Background:
- Traditional Polymerase Chain Reaction (PCR) methods are often time-consuming, expensive, and require extensive sample preparation, including nucleic acid extraction and purification.
- These limitations hinder rapid and widespread diagnostic applications, particularly in resource-limited settings.
Purpose of the Study:
- To develop an ultra-fast and sensitive detection platform for Lumpy Skin Disease Virus (LSDV) that eliminates the need for nucleic acid extraction.
- To improve the speed, sensitivity, and accessibility of LSDV diagnostics for field applications.
Main Methods:
- Utilized a RoomTemp Sample Lysis Kit for rapid nucleic acid release (3 minutes) and inhibition removal.
- Employed ultra-fast Polymerase Chain Reaction (UF-PCR) for 40 cycles in under 16 minutes.
- Integrated CRISPR/Cas12a technology for enhanced sensitivity and detection of LSDV.
- Developed a portable platform providing both fluorescence and Lateral Flow Dipstick (LFD) signals for convenient readout.
Main Results:
- Achieved a detection limit as low as 2 × 101 copies·μL-1 for LSDV.
- Demonstrated a complete detection cycle significantly faster than conventional methods.
- The platform provided visualizable results with excellent sensitivity and high speed.
- Successfully detected Lumpy Skin Disease Virus (LSDV).
Conclusions:
- The developed platform offers a rapid, sensitive, and nucleic acid-extraction-free method for LSDV detection.
- The portable and user-friendly design holds significant potential for field-deployable diagnostics.
- This approach addresses the limitations of traditional PCR, enabling quicker and more accessible disease surveillance.
