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Visualizing in-field detection of HCV using a one-pot RT-RAA-CRISPR/Cas12a platform
Xiangxiang Wei1, Yue Shen1,2, Mingzhu Yuan1
1College of Public Health, Zhengzhou University, No. 100 Kexue Avenue, Zhengzhou 450000, Henan, China. sychen@zzu.edu.cn.
Analytical Methods : Advancing Methods and Applications
|September 23, 2024
Summary
A new Hepatitis C virus RNA (HCV-RNA) diagnostic platform offers rapid, sensitive detection. This visual, one-pot assay uses accessible technology, improving Hepatitis C diagnosis rates in resource-limited settings.
Area of Science:
- Molecular Diagnostics
- Infectious Disease Control
- Biotechnology
Background:
- Hepatitis C poses a significant global health burden, with low diagnostic rates hindering elimination efforts, especially in resource-limited countries.
- Rapid, accurate, and user-friendly point-of-care testing (POCT) assays are crucial for improving Hepatitis C virus (HCV) diagnosis and control.
- Current diagnostic methods may not meet the needs for widespread, accessible screening in all settings.
Purpose of the Study:
- To develop and validate a novel, visual, one-pot Hepatitis C virus RNA (HCV-RNA) detection platform.
- To create an ultra-sensitive, rapid, and user-friendly assay suitable for resource-constrained environments.
- To assess the diagnostic performance of the developed platform using clinical samples.
Main Methods:
- Development of a Visual One-Pot RT-RAA-Cas12a (HCV-VOpRCas12a) platform integrating RT-RAA and CRISPR-Cas12a detection in a single tube.
- Utilization of physically separated reaction components within the tube and low-cost hand warmers for incubation.
- Construction of a visualization device for visual readout of assay results.
Main Results:
- The HCV-VOpRCas12a platform achieved a limit of detection (LoD) as low as 10^0 copies/μL for HCV-RNA.
- Diagnosis of HCV-RNA was accomplished in approximately 15 minutes.
- Clinical validation using 101 serum samples demonstrated 95% sensitivity and 100% specificity.
Conclusions:
- The HCV-VOpRCas12a platform provides a rapid, highly sensitive, and specific method for HCV-RNA detection.
- Its user-friendly design and reliance on accessible components make it suitable for resource-limited settings.
- This platform shows significant potential for advancing the global strategy to eliminate HCV infection and for next-generation molecular diagnostics.
