Rapid diagnosis of duck Tembusu virus and goose astrovirus with TaqMan-based duplex real-time PCR
The mixed infection of duck Tembusu virus (DTMUV) and goose astrovirus (GoAstV) is an important problem that endangers the goose industry. Although quantitative PCR has been widely used in monitoring these two viruses, there is no reliable method to detect them at the same time. In this study, by an...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-03-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1146241/full |
| _version_ | 1797856260752670720 |
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| author | Haiqin Li Haiqin Li Chunhe Wan Zhangzhang Wang Jia Tan Meifang Tan Yanbing Zeng Jiangnan Huang Yu Huang Qi Su Zhaofeng Kang Xiaoquan Guo |
| author_facet | Haiqin Li Haiqin Li Chunhe Wan Zhangzhang Wang Jia Tan Meifang Tan Yanbing Zeng Jiangnan Huang Yu Huang Qi Su Zhaofeng Kang Xiaoquan Guo |
| author_sort | Haiqin Li |
| collection | DOAJ |
| description | The mixed infection of duck Tembusu virus (DTMUV) and goose astrovirus (GoAstV) is an important problem that endangers the goose industry. Although quantitative PCR has been widely used in monitoring these two viruses, there is no reliable method to detect them at the same time. In this study, by analyzing the published genomes of DTMUV and goose astrovirus genotype 2 (GoAstV-2) isolated in China, we found that both viruses have high conservation, showing 96.5 to 99.5% identities within different strains of DTMUV and GoAstV, respectively. Subsequently, PCR primers and TaqMan probes were designed to identify DTMUV and GoAstV-2, and different fluorescent reporters were given to two probes for differential diagnosis. Through the optimization and verification, this study finally developed a duplex TaqMan qPCR method that can simultaneously detect the above two viruses. The lower limits of detection were 100 copies/μL and 10 copies/μL for DTMUV and GoAstV-2 under optimal condition. The assay was also highly specific in detecting one or two viruses in various combinations in specimens, and provide tool for clinical diagnosis of mixed infections of viruses in goose. |
| first_indexed | 2024-04-09T20:37:30Z |
| format | Article |
| id | doaj.art-1a23c33b8ca24d16a3ca4142a50e5beb |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-09T20:37:30Z |
| publishDate | 2023-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-1a23c33b8ca24d16a3ca4142a50e5beb2023-03-30T07:06:41ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2023-03-011410.3389/fmicb.2023.11462411146241Rapid diagnosis of duck Tembusu virus and goose astrovirus with TaqMan-based duplex real-time PCRHaiqin Li0Haiqin Li1Chunhe Wan2Zhangzhang Wang3Jia Tan4Meifang Tan5Yanbing Zeng6Jiangnan Huang7Yu Huang8Qi Su9Zhaofeng Kang10Xiaoquan Guo11Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, ChinaInstitute of Animal Husbandry and Veterinary Medicine, Jiangxi Academy of Agricultural Sciences, Nanchang, Jiangxi, ChinaInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian, ChinaXingguo County Agricultural Technology Extension Center, Ganzhou, Jiangxi, ChinaInstitute of Animal Husbandry and Veterinary Medicine, Jiangxi Academy of Agricultural Sciences, Nanchang, Jiangxi, ChinaInstitute of Animal Husbandry and Veterinary Medicine, Jiangxi Academy of Agricultural Sciences, Nanchang, Jiangxi, ChinaInstitute of Animal Husbandry and Veterinary Medicine, Jiangxi Academy of Agricultural Sciences, Nanchang, Jiangxi, ChinaInstitute of Animal Husbandry and Veterinary Medicine, Jiangxi Academy of Agricultural Sciences, Nanchang, Jiangxi, ChinaInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian, ChinaCollege of Veterinary Medicine, Shandong Agricultural University, Taian, Shandong, ChinaInstitute of Animal Husbandry and Veterinary Medicine, Jiangxi Academy of Agricultural Sciences, Nanchang, Jiangxi, ChinaJiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, ChinaThe mixed infection of duck Tembusu virus (DTMUV) and goose astrovirus (GoAstV) is an important problem that endangers the goose industry. Although quantitative PCR has been widely used in monitoring these two viruses, there is no reliable method to detect them at the same time. In this study, by analyzing the published genomes of DTMUV and goose astrovirus genotype 2 (GoAstV-2) isolated in China, we found that both viruses have high conservation, showing 96.5 to 99.5% identities within different strains of DTMUV and GoAstV, respectively. Subsequently, PCR primers and TaqMan probes were designed to identify DTMUV and GoAstV-2, and different fluorescent reporters were given to two probes for differential diagnosis. Through the optimization and verification, this study finally developed a duplex TaqMan qPCR method that can simultaneously detect the above two viruses. The lower limits of detection were 100 copies/μL and 10 copies/μL for DTMUV and GoAstV-2 under optimal condition. The assay was also highly specific in detecting one or two viruses in various combinations in specimens, and provide tool for clinical diagnosis of mixed infections of viruses in goose.https://www.frontiersin.org/articles/10.3389/fmicb.2023.1146241/fulldiagnosisTembusu virusgoose astrovirusTaqManduplex qPCR |
| spellingShingle | Haiqin Li Haiqin Li Chunhe Wan Zhangzhang Wang Jia Tan Meifang Tan Yanbing Zeng Jiangnan Huang Yu Huang Qi Su Zhaofeng Kang Xiaoquan Guo Rapid diagnosis of duck Tembusu virus and goose astrovirus with TaqMan-based duplex real-time PCR Frontiers in Microbiology diagnosis Tembusu virus goose astrovirus TaqMan duplex qPCR |
| title | Rapid diagnosis of duck Tembusu virus and goose astrovirus with TaqMan-based duplex real-time PCR |
| title_full | Rapid diagnosis of duck Tembusu virus and goose astrovirus with TaqMan-based duplex real-time PCR |
| title_fullStr | Rapid diagnosis of duck Tembusu virus and goose astrovirus with TaqMan-based duplex real-time PCR |
| title_full_unstemmed | Rapid diagnosis of duck Tembusu virus and goose astrovirus with TaqMan-based duplex real-time PCR |
| title_short | Rapid diagnosis of duck Tembusu virus and goose astrovirus with TaqMan-based duplex real-time PCR |
| title_sort | rapid diagnosis of duck tembusu virus and goose astrovirus with taqman based duplex real time pcr |
| topic | diagnosis Tembusu virus goose astrovirus TaqMan duplex qPCR |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1146241/full |
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