Related Experiment Video
Updated: Jun 25, 2026
09:03
Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a
Published on: December 23, 2022
2.8K
Amplification-Free CRISPR/Cas12a-Based Electrochemical Biosensor with Enhanced Sensitivity for Viral Detection.
Yejin Lee1,2, Jin-Ho Lee3,4, Taek Lee5
1Department of Biomedical-Chemical Engineering, The Catholic University of Korea, 43 Jibong-ro, Wonmi-gu, Bucheon-si, Gyeonggi-do 14662, Republic of Korea.
ACS Sensors
|May 22, 2025
Summary
This study introduces a new CRISPR/Cas12a electrochemical biosensor for rapid viral nucleic acid detection. It achieves ultrasensitive, amplification-free detection of human papillomavirus (HPV) at 1 fM, simplifying diagnostics.
Area of Science:
- Biotechnology
- Molecular Diagnostics
- Biosensor Technology
Background:
- Traditional viral nucleic acid detection methods are often complex, time-consuming, and expensive due to amplification steps and specialized probes.
- CRISPR/Cas12a systems show promise for nucleic acid biosensing but require enhanced sensitivity and simplified protocols.
- There is a need for rapid, ultrasensitive, and amplification-free biosensors for early viral detection.
Purpose of the Study:
- To develop a simplified CRISPR/Cas12a-based electrochemical biosensor for ultrasensitive viral nucleic acid detection.
- To introduce novel components, mismatch Ag probe (MAP) and gold electrode on indium tin oxide with a nano array (GELITION), for enhanced biosensing.
- To validate the biosensor's performance for detecting specific viral DNA sequences without amplification.
Main Methods:
- Development of a CRISPR/Cas12a system integrated with a novel mismatch Ag probe (MAP) and a GELITION electrode.
- Electrochemical detection of nucleic acids without requiring target amplification steps.
- Validation using human papillomavirus types 16 and 18 (HPV16 and HPV18) DNA.
Main Results:
- The developed CRISPR/Cas12a electrochemical biosensor achieved amplification-free detection of viral nucleic acids.
- Ultrasensitive detection was demonstrated with a limit of detection (LOD) as low as 1 femtomolar (fM) for HPV16 and HPV18 DNA.
- The biosensor successfully detected target nucleic acids without complex or time-consuming amplification procedures.
Conclusions:
- The novel CRISPR/Cas12a-based electrochemical biosensor offers a simple, rapid, and ultrasensitive platform for nucleic acid detection.
- The combination of MAP and GELITION significantly enhances the sensitivity of CRISPR/Cas12a systems for viral detection.
- This technology holds potential for the early detection of various viruses and could aid in managing future pandemics.
