Related Experiment Video
Updated: Jun 11, 2025
DNA Virus Detection System Based on RPA-CRISPR/Cas12a-SPM and Deep Learning
Published on: May 10, 2024
Establishment of a Lateral Flow Dipstick Detection Method for Influenza A Virus Based on CRISPR/Cas12a System
Xiaoyan Zhao1, Ximing Zheng1, Xiyong Yang1
1Department of Clinical Laboratory, Zhumadian Central Hospital, Zhumadian City, Henan Province, China.
Objective:
This study aimed to develop a rapid, visual PCR-CRISPR/Cas12-LFD method for detecting influenza A by utilizing the conserved region of the matrix protein gene.
Method:
We crafted universal degradation primers and clustered regularly interspaced short palindromic repeats RNA (CRISPR RNA, crRNA) targeting the conserved matrix protein gene of the influenza virus (IFV), integrated with lateral flow dipstick (LFD) technology. This new PCR-CRISPR/Cas12-LFD approach was designed to determine its sensitivity and specificity through the analysis of various clinical samples collected in 2023.
Results:
The developed nucleic acid assay for influenza A viruses (IAV) demonstrated a sensitivity of 10 copies/μL without cross-reactivity with other respiratory pathogens. Evaluation of 82 clinical samples showed high concordance with results from fluorescent Polymerase Chain Reaction (PCR), achieving a kappa value of 0.95.
Conclusion:
A highly sensitive and specific PCR-CRISPR/Cas12-LFD method has been successfully established for the detection of influenza A, offering a robust tool for its diagnosis and aiding in the prevention and control of this virus.
Insights
A new PCR-CRISPR/Cas12-LFD test rapidly detects influenza A virus (IAV) using conserved matrix gene regions. This sensitive and specific method offers a valuable tool for IAV diagnosis and control.
Area of Science:
- Molecular Biology
- Virology
- Biotechnology
Background:
- Influenza A virus (IAV) poses a significant public health threat.
- Accurate and rapid diagnostic tools are crucial for effective management and control of influenza outbreaks.
Purpose of the Study:
- To develop a rapid, visual, and highly sensitive detection method for Influenza A virus.
- To utilize Polymerase Chain Reaction (PCR) combined with CRISPR/Cas12 and Lateral Flow Dipstick (LFD) technology.
Main Methods:
- Designed universal degradation primers and clustered regularly interspaced short palindromic repeats RNA (CRISPR RNA, crRNA) targeting the conserved matrix protein gene of IAV.
- Integrated the CRISPR/Cas12 system with LFD technology for visual detection.
- Validated the assay's sensitivity and specificity using clinical samples.
Main Results:
- The developed PCR-CRISPR/Cas12-LFD assay achieved a sensitivity of 10 copies/μL.
- Demonstrated no cross-reactivity with other common respiratory pathogens.
- Showed high concordance (kappa value of 0.95) with fluorescent PCR when testing 82 clinical samples.
Conclusions:
- Successfully established a highly sensitive and specific PCR-CRISPR/Cas12-LFD method for Influenza A detection.
- The assay provides a robust and rapid tool for IAV diagnosis.
- This method can aid in the prevention and control of influenza virus infections.
