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DNA Virus Detection System Based on RPA-CRISPR/Cas12a-SPM and Deep Learning
Published on: May 10, 2024
818
Rapid, sensitive, and highly specific detection of monkeypox virus by CRISPR-based diagnostic platform
Lin Gong1, Xiaomin Chen1, Yimei Wang1
1Department of Disinfection and Pest Control, Wuhan Center for Disease Control and Prevention, Wuhan, Hubei, China.
Frontiers in Public Health
|July 13, 2023
Summary
A new Monkeypox virus (MPXV) test, MPXV-RCC, combines RPA and CRISPR/Cas12a for rapid detection and clade differentiation. This accurate diagnostic tool can be completed within one hour, aiding global health surveillance.
Area of Science:
- Molecular biology
- Virology
- Diagnostic assay development
Background:
- Monkeypox virus (MPXV) outbreaks necessitate rapid and reliable diagnostic methods.
- Accurate differentiation between MPXV clades (Central Africa and West Africa) is crucial for epidemiological tracking.
Purpose of the Study:
- To develop a novel diagnostic assay for MPXV detection and clade differentiation.
- To evaluate the sensitivity, specificity, and practicality of the developed assay.
Main Methods:
- Combined recombinase polymerase amplification (RPA) with CRISPR/Cas12a-based detection.
- Optimized RPA and CRISPR/Cas12a reaction conditions for MPXV detection.
- Validated the assay using pseudovirus and spiked clinical samples.
Main Results:
- The MPXV-RCC assay achieved detection limits of 5-10 copies/reaction for both MPXV clades.
- The assay accurately differentiated between the Central Africa and West Africa clades.
- The entire diagnostic process, from sample to result, was completed in under one hour.
- No cross-reactivity was observed with other pathogens.
Conclusions:
- The MPXV-RCC assay is a highly sensitive and specific tool for MPXV diagnosis.
- This assay offers a rapid and reliable method for MPXV detection and clade differentiation.
- MPXV-RCC is a promising tool for timely diagnosis and management of MPXV infections.
