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A LAMP-CRISPR/Cas12b rapid detection platform for canine parvovirus detection
Yuting Chen1, Xinyu Zhang1, Gui Hu1
1Hunan Key Laboratory of Biomedical Nanomaterials and Devices, Hunan University of Technology, Zhuzhou, China. huier_88@vip.163.com.
Analytical Methods : Advancing Methods and Applications
|July 25, 2024
Summary
A new LAMP-CRISPR/Cas12b platform offers rapid, sensitive detection of canine parvovirus (CPV). This method is 100 times more sensitive than qPCR, enabling earlier diagnosis and treatment for affected dogs.
Area of Science:
- Veterinary Diagnostics
- Molecular Biology
- Infectious Disease Detection
Background:
- Canine parvovirus (CPV) causes severe diarrhea in dogs, posing a significant threat to the canine industry.
- Timely and accurate CPV detection is crucial for effective treatment and disease control.
Purpose of the Study:
- To develop a novel Loop-mediated isothermal amplification (LAMP) combined with CRISPR-Cas12b platform for rapid CPV detection.
- To establish a sensitive, specific, and efficient diagnostic tool for CPV.
Main Methods:
- Developed a "two-step" and a "one-tube" LAMP-CRISPR/Cas12b assay targeting a conserved CPV gene fragment.
- Utilized specific LAMP primers and single guide RNA (sgRNA) for CRISPR-Cas12b detection.
- Created a point-of-care testing (POCT) platform using magnetic nanoparticle enrichment for nucleic acid extraction and detection.
Main Results:
- The LAMP-CRISPR/Cas12b method achieved a detection limit of 10-1 copies/μL, outperforming qPCR and LAMP by 100-fold sensitivity.
- The assay demonstrated 100% consistency with qPCR for simulated samples and provided results within 1 hour.
- No cross-reactivity was observed with other common canine diarrhea-causing viruses.
Conclusions:
- The developed LAMP-CRISPR/Cas12b platform offers a highly sensitive and specific method for rapid CPV detection.
- This technology provides valuable technical support for early clinical diagnosis and management of CPV infections in dogs.
