Lateral Flow Strip-Compatible Nucleic Acid Testing for Facile Diagnosis of Infectious Pet Diseases

Area of Science:

• Veterinary Diagnostics
• Molecular Biology
• Pathogen Detection

Background:

• Current pet disease diagnostics, like lateral flow immunoassays (LFIA), lack sensitivity, potentially delaying illness identification and treatment.
• Rapid pathogen detection is crucial for timely veterinary care, cost reduction, and preventing zoonotic disease transmission.

Purpose of the Study:

• To develop a rapid, sensitive, and specific diagnostic method for infectious pet diseases.
• To combine recombinase polymerase amplification (RPA) with lateral flow assay (LFA) for improved pathogen detection.

Main Methods:

• Developed a Recombinase Polymerase Amplification (RPA) assay compatible with Lateral Flow Assay (LFA) detection.
• Incorporated T7 exonuclease (T7 Exo) for enhanced specificity of RPA amplicons.
• Validated the assay for feline panleukopenia virus (FPV) and canine parvovirus (CPV) detection in animal samples.

Main Results:

• The proposed RPA-LFA demonstrated a 10,000-fold increase in sensitivity for FPV detection compared to conventional LFIA.
• Results from real cat fecal samples correlated well with PCR-based detection.
• Successfully achieved simultaneous FPV and CPV detection using a dual-line LFA.

Conclusions:

• The RPA-LFA is a highly sensitive and rapid diagnostic tool for infectious pet diseases.
• This technology offers a promising alternative for point-of-care diagnostics in veterinary medicine.
• The method's universality and ability for multiplex detection highlight its potential for broad application.