PED-TRFIA: PEDV early on-site detection based on the time-resolved fluorescence ImmunoAssay

Area of Science:

• Veterinary Diagnostics
• Immunotechnology
• Biosensing

Background:

• Porcine Epidemic Diarrhea Virus (PEDV) poses a significant threat to the global swine industry.
• Current on-site detection methods often lack sufficient sensitivity.
• Existing signal labels like colloidal gold nanoparticles and quantum dots have limitations.

Purpose of the Study:

• To develop a highly sensitive, on-site detection method for PEDV.
• To enable early detection of both PEDV GI and GII genotypes.
• To overcome the sensitivity limitations of current immunoassay labels.

Main Methods:

• Development of a Time-Resolved Fluorescence Immunoassay (PED-TRFIA).
• Utilized time-resolved fluorescence microspheres (TRFMs) with Eu3+ nanoparticles for signal amplification.
• Integrated PED-TRFIA with a paper-based lateral flow immunoassay for field application.

Main Results:

• Achieved a lower limit of detection (LOD) of 9.76 × 10^2 TCID50/mL, 64 times higher sensitivity than commercial kits.
• Demonstrated excellent selectivity against other swine viruses and high accuracy (100%) in clinical samples.
• Showcased good repeatability with coefficients of variation (CVs) of 6.52% for positive and 14.35% for negative samples.
• Enabled complete detection within 15 minutes using low-cost, field-deployable devices.

Conclusions:

• PED-TRFIA offers a sensitive and rapid tool for early on-site detection of PEDV GI and GII.
• The method enhances diagnostic capabilities for PEDV prevention and control in field settings.
• This approach overcomes sensitivity issues and facilitates early detection through signal amplification and background suppression.