A rapid visual detection method for porcine teschovirus through reverse transcription recombinase-aided amplification coupled with lateral flow dipstick
Abstract Background Porcine teschovirus (PTV) is an important enteropathogen, associated with symptoms of polioencephalomyelitis, pneumonia, pericarditis, myocarditis, diarrhea, and reproductive disorders in pigs. Rapid and precise diagnostic methods are essential for managing PTV infections. The st...
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BMC
2024-12-01
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| Online Access: | https://doi.org/10.1186/s12917-024-04442-9 |
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| author | Qiu-Yong Chen Zhi-Hua Sun Ru-Jing Chen Xue-Min Wu Yong-Liang Che Ren-Jie Wu Jing-Li Qiu Bing He Long-Bai Wang Lun-Jiang Zhou |
| author_facet | Qiu-Yong Chen Zhi-Hua Sun Ru-Jing Chen Xue-Min Wu Yong-Liang Che Ren-Jie Wu Jing-Li Qiu Bing He Long-Bai Wang Lun-Jiang Zhou |
| author_sort | Qiu-Yong Chen |
| collection | DOAJ |
| description | Abstract Background Porcine teschovirus (PTV) is an important enteropathogen, associated with symptoms of polioencephalomyelitis, pneumonia, pericarditis, myocarditis, diarrhea, and reproductive disorders in pigs. Rapid and precise diagnostic methods are essential for managing PTV infections. The study introduced a simple, quick, and visual approach for detecting PTV through the use of RT-RAA coupled with LFD. Results The procedures of RT-RAA-LFD for PTV could be carried out with 1.0 μmol/L primer concentration and 2.0 μmol/L probe concentration at 37 °C for 20 min, and the amplification result could be visualized within 5 min through LFD detection. Meanwhile, the assay established in this study showed no interaction with other associated diarrhea viruses, and has high specificity to PTV, with a minimal detection limit of 10 copies/μL and good repeatability. 128 clinical samples suspected of having a PTV infection were tested by RT-PCR and RT-RAA-LFD, respectively. The total diagnostic coincidence rate was 98.44% (126/128) with a Kappa value of 0.96(K ≥ 0.75), demonstrating a high degree of agreement to detect PTV for the two methods. Conclusions The RT-RAA-LFD assay created in this research displayed quick response, specificity, and sensitivity, capable of successfully detecting PTV in less than 25 min, providing an easy-to-use diagnostic instrument for rapid and visual PTV detection, especially suitable for labs and low-resource environments. |
| format | Article |
| id | doaj-art-8d90e5c6b4654d22832ef04f679fc2c7 |
| institution | Kabale University |
| issn | 1746-6148 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj-art-8d90e5c6b4654d22832ef04f679fc2c72025-01-05T12:33:20ZengBMCBMC Veterinary Research1746-61482024-12-012011910.1186/s12917-024-04442-9A rapid visual detection method for porcine teschovirus through reverse transcription recombinase-aided amplification coupled with lateral flow dipstickQiu-Yong Chen0Zhi-Hua Sun1Ru-Jing Chen2Xue-Min Wu3Yong-Liang Che4Ren-Jie Wu5Jing-Li Qiu6Bing He7Long-Bai Wang8Lun-Jiang Zhou9Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture SciencesInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture SciencesInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture SciencesInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture SciencesInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture SciencesInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture SciencesInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture SciencesInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture SciencesInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture SciencesInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agriculture SciencesAbstract Background Porcine teschovirus (PTV) is an important enteropathogen, associated with symptoms of polioencephalomyelitis, pneumonia, pericarditis, myocarditis, diarrhea, and reproductive disorders in pigs. Rapid and precise diagnostic methods are essential for managing PTV infections. The study introduced a simple, quick, and visual approach for detecting PTV through the use of RT-RAA coupled with LFD. Results The procedures of RT-RAA-LFD for PTV could be carried out with 1.0 μmol/L primer concentration and 2.0 μmol/L probe concentration at 37 °C for 20 min, and the amplification result could be visualized within 5 min through LFD detection. Meanwhile, the assay established in this study showed no interaction with other associated diarrhea viruses, and has high specificity to PTV, with a minimal detection limit of 10 copies/μL and good repeatability. 128 clinical samples suspected of having a PTV infection were tested by RT-PCR and RT-RAA-LFD, respectively. The total diagnostic coincidence rate was 98.44% (126/128) with a Kappa value of 0.96(K ≥ 0.75), demonstrating a high degree of agreement to detect PTV for the two methods. Conclusions The RT-RAA-LFD assay created in this research displayed quick response, specificity, and sensitivity, capable of successfully detecting PTV in less than 25 min, providing an easy-to-use diagnostic instrument for rapid and visual PTV detection, especially suitable for labs and low-resource environments.https://doi.org/10.1186/s12917-024-04442-9Porcine teschovirusVisual detection methodRecombinase-aided amplificationLateral flow dipstick |
| spellingShingle | Qiu-Yong Chen Zhi-Hua Sun Ru-Jing Chen Xue-Min Wu Yong-Liang Che Ren-Jie Wu Jing-Li Qiu Bing He Long-Bai Wang Lun-Jiang Zhou A rapid visual detection method for porcine teschovirus through reverse transcription recombinase-aided amplification coupled with lateral flow dipstick BMC Veterinary Research Porcine teschovirus Visual detection method Recombinase-aided amplification Lateral flow dipstick |
| title | A rapid visual detection method for porcine teschovirus through reverse transcription recombinase-aided amplification coupled with lateral flow dipstick |
| title_full | A rapid visual detection method for porcine teschovirus through reverse transcription recombinase-aided amplification coupled with lateral flow dipstick |
| title_fullStr | A rapid visual detection method for porcine teschovirus through reverse transcription recombinase-aided amplification coupled with lateral flow dipstick |
| title_full_unstemmed | A rapid visual detection method for porcine teschovirus through reverse transcription recombinase-aided amplification coupled with lateral flow dipstick |
| title_short | A rapid visual detection method for porcine teschovirus through reverse transcription recombinase-aided amplification coupled with lateral flow dipstick |
| title_sort | rapid visual detection method for porcine teschovirus through reverse transcription recombinase aided amplification coupled with lateral flow dipstick |
| topic | Porcine teschovirus Visual detection method Recombinase-aided amplification Lateral flow dipstick |
| url | https://doi.org/10.1186/s12917-024-04442-9 |
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