Rapid Detection of Fish Viruses Using RPA-CRISPR/Cas12a
Author Information
Author(s): Lu Zhoutao, Liang Mincong, Li Chuanrui, Xu Yichun, Weng Shaoping, He Jianguo, Guo Changjun
Primary Institution: School of Marine Sciences, State Key Laboratory for Biocontrol, Southern Marine Science and Engineering Guangdong Laboratory, Sun Yat-sen University, Guangzhou, China
Hypothesis
Can a rapid, sensitive, and visual detection method for mandarin fish ranavirus and infectious spleen and kidney necrosis virus be developed using the RPA-CRISPR/Cas12a system?
Conclusion
A new detection method for ISKNV and MRV has been established, providing a powerful tool for the prompt diagnosis of these aquatic pathogens.
Supporting Evidence
- The method can detect as low as 1 copy/μL of MRV and 0.1 copy/μL of ISKNV.
- The detection can be performed at a constant temperature of 37–39°C.
- A 30-min RPA amplification followed by a 15-min CRISPR/Cas reaction is sufficient for detecting most samples.
- Two probes were identified that can be observed without the need for excitation light.
Takeaway
Scientists created a quick and easy way to find harmful viruses in fish, which helps keep fish healthy and saves money for fish farmers.
Methodology
The study used a recombinase polymerase amplification (RPA)−CRISPR/Cas12a system to detect MRV and ISKNV.
Limitations
Nucleic acid extraction and purification from samples are required for detection.
Statistical Information
P-Value
p<0.05
Statistical Significance
p<0.05
Digital Object Identifier (DOI)
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