Related Experiment Video
Updated: Jun 25, 2025
Author Spotlight: Advancements and Challenges in Hepatitis B Virus Detection
Published on: December 15, 2023
CRISPR/Cas12b assisted loop-mediated isothermal amplification for easy, rapid and sensitive quantification of chronic
Haipo Xu1, Gengping Lin2, Ronghua Chen3
1The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, 350025, PR China; Mengchao Med-X Center, Fuzhou University, Fuzhou, 350116, PR China; College of Chemical Engineering, Fuzhou University, Fuzhou, 350116, PR China.
A new CRISPR/Cas12b-assisted assay offers rapid, sensitive detection of hepatitis B virus (HBV) DNA. This point-of-care test is ideal for screening chronic hepatitis B (CHB) in resource-limited settings.
Area of Science:
- Molecular diagnostics
- Biotechnology
- Infectious disease detection
Background:
- Chronic hepatitis B (CHB) causes high mortality, with millions undiagnosed annually.
- Existing PCR-based assays are not suitable for high-burden, resource-limited settings.
- CRISPR/Cas systems offer sensitive and specific point-of-care testing (POCT) potential.
Purpose of the Study:
- To develop a rapid, sensitive, and precise method for hepatitis B virus (HBV) DNA quantification.
- To create a point-of-care diagnostic assay suitable for resource-limited settings.
- To address the urgent need for effective HBV detection methods.
Main Methods:
- A one-pot, one-step CRISPR/Cas12b assisted loop-mediated isothermal amplification (LAMP) assay was developed.
- Optimization of reaction buffer, enzyme concentration, reporter, temperature, and additives.
- Validation using 236 clinical samples compared to real-time quantitative PCR.
Main Results:
- The assay quantifies HBV DNA within 1 hour with a limit of detection of 25 copies/mL.
- Demonstrated 99.00% sensitivity and 100.00% specificity in clinical samples.
- Showed no cross-reactivity with other blood-borne pathogens.
Conclusions:
- The developed CRISPR/Cas12b-LAMP assay is a convenient and reliable point-of-care diagnostic tool.
- It significantly improves HBV diagnosis capabilities, especially in resource-limited areas.
- The assay is advantageous for rapid CHB screening and evaluating therapeutic efficacy.
