Development of a Portable Rapid Detection Method for Porcine Epidemic Diarrhea Virus Using Reverse Transcription-Recombinase-Aided Amplification Technology
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Joint International Research Laboratory of Animal Health and Food Safety of Ministry of Education, Single Molecule Biochemistry & Biomedicine Laboratory (Sinmolab), College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
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Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
*
Author to whom correspondence should be addressed.
Animals 2025, 15(2), 281; https://doi.org/10.3390/ani15020281
Submission received: 10 December 2024
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Revised: 15 January 2025
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Accepted: 17 January 2025
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Published: 20 January 2025
(This article belongs to the Special Issue Infection and Immunity Response Mechanism of Coronavirus in Animals)
Simple Summary
Porcine epidemic diarrhea virus (PEDV), a member of the Coronaviridae family, is responsible for severe watery diarrhea, vomiting, and dehydration, leading to significant mortality in neonatal piglets. A rapid detection system for PEDV, based on reverse transcription-recombinase-aided amplification (RT-RAA) technology, was successfully developed in this study. The developed detection system is characterized by its simplicity in operation, high sensitivity, and stability. Additionally, when integrated with a compact blue-light detection device, it facilitates on-site visual detection, making it suitable for field applications.
Abstract
Porcine epidemic diarrhea virus (PEDV) continues to spread globally, causing clinical symptoms in piglets, including watery diarrhea, vomiting, and dehydration. Its exceptionally high morbidity and mortality rate contributes significantly to the economic losses of the swine industry. The continuous genetic mutations of PEDV have compromised the effectiveness of classical strain vaccines. Early detection and accurate diagnosis are therefore crucial for controlling its further spread. Developing a detection method that is user-friendly, highly sensitive, and efficient is crucial for disease control. In this study, a point-of-care rapid detection method for PEDV was successfully established using reverse transcription-recombinase-aided amplification (RT-RAA) technology. This method enables results to be obtained within 20 min of amplification at a constant temperature of 42 °C. It demonstrates high sensitivity, with a detection limit as low as 1 copy/μL, and shows strong specificity, with no cross-reactivity observed with seven other common swine pathogens. When applied to clinical samples, the results were 100% consistent with those obtained by RT-qPCR. This method is distinguished by its portable instrumentation and simple operation, making it particularly suitable for resource-constrained settings.
