Journal of virological methodsMar 24, 2026

Fast, extraction-free test using LAMP and CRISPR to detect pseudorabies virus

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Mar 25, 2026

Abstract

The LAMP-CRISPR/Cas12a detection system achieved a detection limit of 1.0×10⁻⁴ copies/μL for porcine pseudorabies virus (PRV).

The system demonstrated superior sensitivity compared to PCR-CRISPR/Cas12a and qPCR methods.
Specificity testing confirmed the detection of PRV without cross-reactivity to other porcine pathogens.
A comparison of 26 serum samples showed 100% agreement in positive results between the LAMP-CRISPR/Cas12a and PCR-CRISPR/Cas12a systems.
Both methods detected 12 positive samples in the comparison.
Results are available within one hour and can be interpreted visually without specialized equipment.

Simplified

This study developed a LAMP-CRISPR/Cas12a detection system for rapid and visual identification of porcine pseudorabies virus (PRV). Optimal sgRNA and LAMP-specific primers were designed based on the conserved sequences of the viral pathogenic gene gG. The combined detection system demonstrated superior sensitivity compared to PCR-CRISPR/Cas12a and qPCR methods, achieving a detection limit of 1.0 × 10⁻⁴ copies/μL for the target plasmid DNA. Specificity testing confirmed the selective identification of PRV without cross-reactivity to other porcine pathogens. Parallel comparison of 26 serum samples between LAMP-CRISPR/Cas12a and PCR-CRISPR/Cas12a systems showed 100% concordance for positive results, with both detecting 12 positive samples. The method eliminates the need for viral nucleic acid extraction and requires only a constant temperature device and/or basic fluorescence detection equipment. Results are obtainable within one hour and are readable by the naked eye. This simple, sensitive, and equipment-independent approach is ideal for on-site rapid diagnosis of pseudorabies in pigs, offering significant applications in clinical diagnosis, epidemiological surveillance, and field testing.

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Competing interests

Declaration of Competing Interest The authors declare that there are no conflicts of interest regarding the publication of this paper.

PubMed

Fast, extraction-free test using LAMP and CRISPR to detect pseudorabies virus

Abstract

Full Text

Funding

Competing interests

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