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CRISPR/Cas12a-based MUSCA-PEC strategy for HSV-1 assay
Bohao Dou1, Yinhao Zhang1, Huiyu Gao1
1Key Laboratory of Optic-electric Sensing and Analytical Chemistry for Life Science, MOE, Shandong Key Laboratory of Biochemical Analysis, Key Laboratory of Analytical Chemistry for Life Science in Universities of Shandong, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao, 266042, China.
A new photoelectrochemical method detects Herpes simplex virus (HSV-1) using CRISPR/Cas12a and a novel signal amplification technique. This approach offers direct, fast, and ultrasensitive detection of HSV-1 in human serum.
Area of Science:
- Biotechnology
- Biosensing
- Molecular Diagnostics
Background:
- Photoelectrochemical (PEC) sensing typically uses constant potential excitation, necessitating novel signal acquisition methods.
- Developing sensitive and rapid detection strategies for viruses like Herpes simplex virus (HSV-1) is crucial for clinical diagnostics.
Purpose of the Study:
- To develop a novel photoelectrochemical strategy for ultrasensitive Herpes simplex virus type 1 (HSV-1) detection.
- To integrate CRISPR/Cas12a technology with a multiple potential step chronoamperometry (MUSCA) pattern for enhanced signal amplification.
Main Methods:
- A CRISPR/Cas12a system was employed, triggered by entropy-driven target recycling for HSV-1 detection.
- Multiple potential step chronoamperometry (MUSCA) was utilized as a signal amplifier to enhance photocurrent responses.
- The strategy involved Cas12a activation, cleavage of a circular fragment, release of alkaline phosphatase (ALP), and subsequent catalysis of p-Aminophenol (p-AP).
Main Results:
- The developed strategy achieved a superior detection limit of 3 aM for HSV-1.
- The MUSCA technique provided direct, fast, and ultrasensitive signal enhancement.
- The method demonstrated successful application in detecting HSV-1 in human serum samples.
Conclusions:
- The combination of MUSCA and CRISPR/Cas12a offers a powerful platform for nucleic acid detection.
- This novel approach presents a promising advancement for direct, rapid, and ultrasensitive viral detection in clinical settings.
