Development of a Portable Rapid Detection Method for Porcine Epidemic Diarrhea Virus Using Reverse Transcription-Recombinase-Aided Amplification Technology
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...
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MDPI AG
2025-01-01
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| Series: | Animals |
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| author | Yiran Zhao Weijie Yi Qicheng Yang Jiahao Li Yanke Shan Fei Liu |
| author_facet | Yiran Zhao Weijie Yi Qicheng Yang Jiahao Li Yanke Shan Fei Liu |
| author_sort | Yiran Zhao |
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| description | 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. |
| format | Article |
| id | doaj-art-b03309a3fcdc4467a2083c82cb68d17a |
| institution | Kabale University |
| issn | 2076-2615 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
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| series | Animals |
| spelling | doaj-art-b03309a3fcdc4467a2083c82cb68d17a2025-01-24T13:18:25ZengMDPI AGAnimals2076-26152025-01-0115228110.3390/ani15020281Development of a Portable Rapid Detection Method for Porcine Epidemic Diarrhea Virus Using Reverse Transcription-Recombinase-Aided Amplification TechnologyYiran Zhao0Weijie Yi1Qicheng Yang2Jiahao Li3Yanke Shan4Fei Liu5Joint 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, ChinaJoint 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, ChinaJoint 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, ChinaKey Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaJoint 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, ChinaJoint 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, ChinaPorcine 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.https://www.mdpi.com/2076-2615/15/2/281porcine epidemic diarrhea virusisothermal amplification technologyreverse transcription-recombinase-aided amplificationpoint-of-care detection |
| spellingShingle | Yiran Zhao Weijie Yi Qicheng Yang Jiahao Li Yanke Shan Fei Liu Development of a Portable Rapid Detection Method for Porcine Epidemic Diarrhea Virus Using Reverse Transcription-Recombinase-Aided Amplification Technology Animals porcine epidemic diarrhea virus isothermal amplification technology reverse transcription-recombinase-aided amplification point-of-care detection |
| title | Development of a Portable Rapid Detection Method for Porcine Epidemic Diarrhea Virus Using Reverse Transcription-Recombinase-Aided Amplification Technology |
| title_full | Development of a Portable Rapid Detection Method for Porcine Epidemic Diarrhea Virus Using Reverse Transcription-Recombinase-Aided Amplification Technology |
| title_fullStr | Development of a Portable Rapid Detection Method for Porcine Epidemic Diarrhea Virus Using Reverse Transcription-Recombinase-Aided Amplification Technology |
| title_full_unstemmed | Development of a Portable Rapid Detection Method for Porcine Epidemic Diarrhea Virus Using Reverse Transcription-Recombinase-Aided Amplification Technology |
| title_short | Development of a Portable Rapid Detection Method for Porcine Epidemic Diarrhea Virus Using Reverse Transcription-Recombinase-Aided Amplification Technology |
| title_sort | development of a portable rapid detection method for porcine epidemic diarrhea virus using reverse transcription recombinase aided amplification technology |
| topic | porcine epidemic diarrhea virus isothermal amplification technology reverse transcription-recombinase-aided amplification point-of-care detection |
| url | https://www.mdpi.com/2076-2615/15/2/281 |
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