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Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases
Published on: August 16, 2024
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A CRISPR Test for Detection of Circulating Nuclei Acids.
Jen-Hui Tsou1, Qixin Leng1, Feng Jiang1
1Department of Pathology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD, USA.
Translational Oncology
|October 22, 2019
Summary
A new CRISPR-Cas12a system enables rapid, point-of-care detection of human papillomavirus (HPV) DNA directly in plasma. This innovation simplifies molecular diagnostics for cervical cancer, requiring no complex equipment for visual results.
Area of Science:
- Molecular Biology
- Biotechnology
- Diagnostics
Background:
- CRISPR-Cas12a offers sensitive nucleic acid detection for disease diagnosis.
- Current CRISPR-Cas12a systems require specialized equipment, limiting point-of-care (POC) applications.
- Human papillomavirus (HPV) DNA detection is crucial for cervical cancer diagnosis.
Purpose of the Study:
- To develop a CRISPR-Cas12a-based POC test for direct detection of circulating HPV DNA in plasma.
- To enable rapid, visual readout of HPV DNA detection without complex instrumentation.
- To assess the system's sensitivity and specificity for HPV16 and HPV18 detection.
Main Methods:
- CRISPR-Cas12a system integrated with lateral-flow strips.
- Isothermal amplification of target DNA from plasma without prior isolation.
- Incubation with Cas12a, crRNA, and fluorescent-biotin reporters for detection.
Main Results:
- Direct and specific detection of HPV16 and HPV18 DNA in liquid samples.
- Achieved a limit of detection of 0.24 fM, comparable to PCR but faster.
- Successfully detected HPV16 and HPV18 in plasma samples from cervical cancer patients.
Conclusions:
- A novel CRISPR-Cas12a POC system was developed for convenient detection of circulating HPV DNA.
- The system eliminates the need for technical expertise and ancillary machinery.
- This technology holds promise for accessible molecular diagnostics in resource-limited settings.
