Multiplex one-step RT‒qPCR assays for simultaneous detection of AMDV, MEV and CDV
Abstract Background Aleutian mink disease, mink viral enteritis and canine distemper are known as the three most serious diseases that cause great economic loss in the mink industry. In clinical practice, aleutian mink disease virus (AMDV), mink enteritis virus (MEV) and canine distemper virus (CDV)...
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BMC
2025-01-01
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| Series: | BMC Veterinary Research |
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| Online Access: | https://doi.org/10.1186/s12917-024-04349-5 |
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| author | Zhi Cao Hang Xu Xinru Zhao Ke Zhang Dehua Yin Shuai Ma Wenling Li Siyu Li Jianwei Ren Jianxin Wen |
| author_facet | Zhi Cao Hang Xu Xinru Zhao Ke Zhang Dehua Yin Shuai Ma Wenling Li Siyu Li Jianwei Ren Jianxin Wen |
| author_sort | Zhi Cao |
| collection | DOAJ |
| description | Abstract Background Aleutian mink disease, mink viral enteritis and canine distemper are known as the three most serious diseases that cause great economic loss in the mink industry. In clinical practice, aleutian mink disease virus (AMDV), mink enteritis virus (MEV) and canine distemper virus (CDV) are common mixed infections, and they have similar clinical clinical signs, such as diarrhoea. Therefore, a rapid and accurate differential diagnosis method for use on mink ranches is essential for the control of these three pathogens. Here, we developed multiplex one-step real-time quantitative PCR (RT‒qPCR) assays for the simultaneous detection and quantification of AMDV, MEV and CDV by using three primers and probes based on the conserved NS1, VP2 and N genes, respectively. Results The results showed that the established method can not cross-react with other mink pathogens, with a detection sensitivity of 25 copies/µL and a coefficient of variation less than 3.51%. Moreover, the interference experiment showed that the presence of AMDV, MEV and CDV templates at different concentrations would not interfere with the detection results. Furthermore, two hundred clinical samples of mink with diarrhoea were simultaneously analysed using multiplex RT‒qPCR and single RT‒qPCR, the Kappa values were all greater than 0.921, indicating that there was a high degree of coincidence between the two detection methods. Conclusions In conclusion, multiplex RT‒qPCR exhibited high specificity, sensitivity, and reproducibility, indicating that this method can be used as a reliable and specific tool for the differential detection and quantification of AMDV, MEV and CDV. |
| format | Article |
| id | doaj-art-7e1723d32cfb4efbba0fcbd7ae5efa45 |
| institution | Kabale University |
| issn | 1746-6148 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj-art-7e1723d32cfb4efbba0fcbd7ae5efa452025-01-19T12:27:16ZengBMCBMC Veterinary Research1746-61482025-01-012111910.1186/s12917-024-04349-5Multiplex one-step RT‒qPCR assays for simultaneous detection of AMDV, MEV and CDVZhi Cao0Hang Xu1Xinru Zhao2Ke Zhang3Dehua Yin4Shuai Ma5Wenling Li6Siyu Li7Jianwei Ren8Jianxin Wen9College of Veterinary Medicine, Qingdao Agricultural UniversityCollege of Veterinary Medicine, Qingdao Agricultural UniversityCollege of Veterinary Medicine, Qingdao Agricultural UniversityCollege of Veterinary Medicine, Qingdao Agricultural UniversityCollege of Veterinary Medicine, Qingdao Agricultural UniversityQingdao Animal Disease Prevention and Control CenterCollege of Veterinary Medicine, Qingdao Agricultural UniversityCollege of Veterinary Medicine, Qingdao Agricultural UniversityCollege of Veterinary Medicine, Qingdao Agricultural UniversityCollege of Veterinary Medicine, Qingdao Agricultural UniversityAbstract Background Aleutian mink disease, mink viral enteritis and canine distemper are known as the three most serious diseases that cause great economic loss in the mink industry. In clinical practice, aleutian mink disease virus (AMDV), mink enteritis virus (MEV) and canine distemper virus (CDV) are common mixed infections, and they have similar clinical clinical signs, such as diarrhoea. Therefore, a rapid and accurate differential diagnosis method for use on mink ranches is essential for the control of these three pathogens. Here, we developed multiplex one-step real-time quantitative PCR (RT‒qPCR) assays for the simultaneous detection and quantification of AMDV, MEV and CDV by using three primers and probes based on the conserved NS1, VP2 and N genes, respectively. Results The results showed that the established method can not cross-react with other mink pathogens, with a detection sensitivity of 25 copies/µL and a coefficient of variation less than 3.51%. Moreover, the interference experiment showed that the presence of AMDV, MEV and CDV templates at different concentrations would not interfere with the detection results. Furthermore, two hundred clinical samples of mink with diarrhoea were simultaneously analysed using multiplex RT‒qPCR and single RT‒qPCR, the Kappa values were all greater than 0.921, indicating that there was a high degree of coincidence between the two detection methods. Conclusions In conclusion, multiplex RT‒qPCR exhibited high specificity, sensitivity, and reproducibility, indicating that this method can be used as a reliable and specific tool for the differential detection and quantification of AMDV, MEV and CDV.https://doi.org/10.1186/s12917-024-04349-5AMDVMEVCDVMultiplex RT‒qPCRDifferential detection |
| spellingShingle | Zhi Cao Hang Xu Xinru Zhao Ke Zhang Dehua Yin Shuai Ma Wenling Li Siyu Li Jianwei Ren Jianxin Wen Multiplex one-step RT‒qPCR assays for simultaneous detection of AMDV, MEV and CDV BMC Veterinary Research AMDV MEV CDV Multiplex RT‒qPCR Differential detection |
| title | Multiplex one-step RT‒qPCR assays for simultaneous detection of AMDV, MEV and CDV |
| title_full | Multiplex one-step RT‒qPCR assays for simultaneous detection of AMDV, MEV and CDV |
| title_fullStr | Multiplex one-step RT‒qPCR assays for simultaneous detection of AMDV, MEV and CDV |
| title_full_unstemmed | Multiplex one-step RT‒qPCR assays for simultaneous detection of AMDV, MEV and CDV |
| title_short | Multiplex one-step RT‒qPCR assays for simultaneous detection of AMDV, MEV and CDV |
| title_sort | multiplex one step rt qpcr assays for simultaneous detection of amdv mev and cdv |
| topic | AMDV MEV CDV Multiplex RT‒qPCR Differential detection |
| url | https://doi.org/10.1186/s12917-024-04349-5 |
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