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Development of a point-of-care diagnostic method for FMDV SAT2 using RT-RAA-CRISPR technology.

Yabo Yuan1, Bo Ni2, Xiaoxiao Tian2

  • 1College of Veterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang Province, China; China Animal Health and Epidemiology Center, Qingdao, Shandong Province, China; National Foot-and-Mouth Disease Professional Laboratory, Qingdao, Shandong Province, China.

International Journal of Biological Macromolecules
|April 30, 2026
PubMed
Summary

A new rapid detection method for foot-and-mouth disease virus (FMDV) SAT2 has been developed using RT-RAA-CRISPR/Cas13a technology. This sensitive and specific tool aids in early FMDV SAT2 diagnosis and control.

Keywords:
CRISPR/Cas13aFMDVLFSOn-site detectionRT–RAASAT2

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Area of Science:

  • Veterinary Virology
  • Molecular Diagnostics
  • Epidemiology

Background:

  • Foot-and-mouth disease virus (FMDV) SAT2 is expanding globally, posing a significant threat to livestock in Africa, the Middle East, and potentially Asia.
  • Current diagnostic methods may lack the speed or specificity required for effective early detection and control of FMDV SAT2 outbreaks.

Purpose of the Study:

  • To develop a novel, highly sensitive, and specific nucleic acid detection method for FMDV SAT2.
  • To create a rapid, field-deployable diagnostic tool for early FMDV SAT2 identification.

Main Methods:

  • Design and optimization of reverse transcription-recombinase-aided amplification (RT-RAA) primers targeting conserved FMDV SAT2 sequences.
  • Integration of CRISPR RNA (crRNA) with the CRISPR/Cas13a system for enhanced detection sensitivity and specificity.
  • Validation using fluorescence readers and lateral flow test strips (LFSs), comparing results with RT-qPCR.

Main Results:

  • A novel RT-RAA-CRISPR/Cas13a based detection method for FMDV SAT2 was successfully established.
  • The method demonstrated high sensitivity, specificity, and no cross-reactivity with other FMDV serotypes or common livestock pathogens.
  • Achieved 100% concordance with RT-qPCR in simulated clinical samples, with easily interpretable results.

Conclusions:

  • The developed RT-RAA-CRISPR/Cas13a method offers a sensitive, specific, and rapid diagnostic tool for FMDV SAT2.
  • Its ease of use, low equipment requirement, and suitability for onsite detection make it valuable for early FMDV SAT2 epidemic control.