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Related Experiment Video

Updated: Jul 28, 2025

Visual Detection of Multiple Nucleic Acids in a Capillary Array
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Multiple accurate and sensitive arrays for Capripoxvirus (CaPV) differentiation.

Gaihua Cao1, Yifan Xiong1, Meimei Shi2

  • 1Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, 400044, PR China.

Analytica Chimica Acta
|May 31, 2023
PubMed
Summary
This summary is machine-generated.

A new CRISPR/Cas12a array combined with Multiple-recombinase polymerase amplification (M-RPA) enables accurate differentiation of Capripoxvirus (CaPV) strains. This method offers sensitive, portable detection for livestock disease control.

Keywords:
CRISPR/Cas12aGoatpox virus (GTPV)Lumpy skin disease virus (LSDV)Multiple-recombinase polymerase amplification (M-RPA)Real-time quantitative PCR (qPCR)Sheeppox virus (SPPV)

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Area of Science:

  • Molecular Biology
  • Veterinary Virology
  • Biotechnology

Background:

  • Capripoxvirus (CaPV) causes significant economic losses in livestock and dairy industries.
  • Distinguishing between the three CaPV types is challenging due to 97% genetic homology.
  • Accurate differentiation is crucial for effective outbreak control and disease management.

Purpose of the Study:

  • To develop a sensitive and accurate method for differentiating Capripoxvirus (CaPV) strains.
  • To establish a rapid, point-of-care diagnostic tool for CaPV detection.
  • To address the technical gap in quantitative PCR (qPCR) for CaPV differentiation.

Main Methods:

  • Development of a CRISPR/Cas12a array integrated with Multiple-recombinase polymerase amplification (M-RPA).
  • Targeting of VARV B22R and RPO30 genes for specific CaPV differentiation.
  • Construction of a Lateral Flow Dipstick (LFD) array for visual detection.
  • Development of a real-time quantitative PCR (qPCR) array for quantitative analysis.

Main Results:

  • The CRISPR/Cas12a-M-RPA assay achieved detection limits as low as 50, 40, and 60 copies for the three CaPV types.
  • The LFD array provided portable and intuitive detection, suitable for point-of-care applications.
  • A qPCR array was successfully constructed to complement existing diagnostic capabilities.

Conclusions:

  • The developed CRISPR/Cas12a array and LFD array offer a practical and effective solution for Capripoxvirus differentiation.
  • This technology enhances diagnostic capabilities for livestock disease surveillance and control.
  • The study provides valuable tools for quarantine departments and veterinary diagnostics.