Rapid detection of goose astrovirus genotypes 2 using real-time reverse transcription recombinase polymerase amplification
Abstract Background Goose astrovirus (GoAstV) is an important pathogen that causes joint and visceral gout in goslings. It has been circulating in many provinces of China since 2017. Goose astrovirus genotypes 2 (GoAstV-2) is the main epidemic strain, and its high morbidity and mortality have caused...
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| Format: | Article |
| Language: | English |
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BMC
2023-11-01
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| Series: | BMC Veterinary Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12917-023-03790-2 |
| _version_ | 1827633884386295808 |
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| author | Haiqin Li Yujun Zhu Chunhe Wan Zhangzhang Wang Lei Liu Meifang Tan Fanfan Zhang Yanbing Zeng Jiangnan Huang Chengcheng Wu Yu Huang Zhaofeng Kang Xiaoqiao Guo |
| author_facet | Haiqin Li Yujun Zhu Chunhe Wan Zhangzhang Wang Lei Liu Meifang Tan Fanfan Zhang Yanbing Zeng Jiangnan Huang Chengcheng Wu Yu Huang Zhaofeng Kang Xiaoqiao Guo |
| author_sort | Haiqin Li |
| collection | DOAJ |
| description | Abstract Background Goose astrovirus (GoAstV) is an important pathogen that causes joint and visceral gout in goslings. It has been circulating in many provinces of China since 2017. Goose astrovirus genotypes 2 (GoAstV-2) is the main epidemic strain, and its high morbidity and mortality have caused huge economic losses to the goose industry. An accurate point-of-care detection for GoAstV-2 is of great significance. In this study, we developed a real-time reverse transcription recombinase polymerase amplification (RT-RPA) method for the on-site detection of GoAstV-2 infection. Results The real-time RT-RPA reaction was carried out at a constant temperature of 39 °C, and the entire detection time from nucleic acid preparation to the end of amplification was only 25 min using the portable device. The results of a specificity analysis showed that no cross-reaction was observed with other related pathogens. The detection limit of the assay was 100 RNA copies/μL. The low coefficient of variation value indicated excellent repeatability. We used 270 clinical samples to evaluate the performance of our established method, the positive concordance rates with RT-qPCR were 99.6%, and the linear regression analysis revealed a strong correlation. Conclusions The established real-time RT-RPA assay showed high rapidity, specificity and sensitivity, which can be widely applied in the laboratory, field and especially in the resource-limited settings for GoAstV-2 point-of-care diagnosis. |
| first_indexed | 2024-03-09T15:05:44Z |
| format | Article |
| id | doaj.art-39a84205222b4755b48e182d5e36db06 |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-03-09T15:05:44Z |
| publishDate | 2023-11-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj.art-39a84205222b4755b48e182d5e36db062023-11-26T13:38:20ZengBMCBMC Veterinary Research1746-61482023-11-011911910.1186/s12917-023-03790-2Rapid detection of goose astrovirus genotypes 2 using real-time reverse transcription recombinase polymerase amplificationHaiqin Li0Yujun Zhu1Chunhe Wan2Zhangzhang Wang3Lei Liu4Meifang Tan5Fanfan Zhang6Yanbing Zeng7Jiangnan Huang8Chengcheng Wu9Yu Huang10Zhaofeng Kang11Xiaoqiao Guo12Institute of Animal Husbandry and Veterinary Medicine, Jiangxi Academy of Agricultural SciencesGuangdong laboratory animals monitoring instituteand Guangdong Provincial Key Laboratory of Laboratory AnimalsInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural SciencesXingguo County Agricultural Technology Extension CenterXinyuYushui District Center for Agricultural SciencesInstitute of Animal Husbandry and Veterinary Medicine, Jiangxi Academy of Agricultural SciencesInstitute of Animal Husbandry and Veterinary Medicine, Jiangxi Academy of Agricultural SciencesInstitute of Animal Husbandry and Veterinary Medicine, Jiangxi Academy of Agricultural SciencesInstitute of Animal Husbandry and Veterinary Medicine, Jiangxi Academy of Agricultural SciencesInstitute of Animal Husbandry and Veterinary Medicine, Jiangxi Academy of Agricultural SciencesInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural SciencesInstitute of Animal Husbandry and Veterinary Medicine, Jiangxi Academy of Agricultural SciencesJiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi AgriculturalUniversityAbstract Background Goose astrovirus (GoAstV) is an important pathogen that causes joint and visceral gout in goslings. It has been circulating in many provinces of China since 2017. Goose astrovirus genotypes 2 (GoAstV-2) is the main epidemic strain, and its high morbidity and mortality have caused huge economic losses to the goose industry. An accurate point-of-care detection for GoAstV-2 is of great significance. In this study, we developed a real-time reverse transcription recombinase polymerase amplification (RT-RPA) method for the on-site detection of GoAstV-2 infection. Results The real-time RT-RPA reaction was carried out at a constant temperature of 39 °C, and the entire detection time from nucleic acid preparation to the end of amplification was only 25 min using the portable device. The results of a specificity analysis showed that no cross-reaction was observed with other related pathogens. The detection limit of the assay was 100 RNA copies/μL. The low coefficient of variation value indicated excellent repeatability. We used 270 clinical samples to evaluate the performance of our established method, the positive concordance rates with RT-qPCR were 99.6%, and the linear regression analysis revealed a strong correlation. Conclusions The established real-time RT-RPA assay showed high rapidity, specificity and sensitivity, which can be widely applied in the laboratory, field and especially in the resource-limited settings for GoAstV-2 point-of-care diagnosis.https://doi.org/10.1186/s12917-023-03790-2Goose astrovirus genotypes 2Real-timeReverse transcription recombinase polymerase amplificationPoint-of-care diagnosis |
| spellingShingle | Haiqin Li Yujun Zhu Chunhe Wan Zhangzhang Wang Lei Liu Meifang Tan Fanfan Zhang Yanbing Zeng Jiangnan Huang Chengcheng Wu Yu Huang Zhaofeng Kang Xiaoqiao Guo Rapid detection of goose astrovirus genotypes 2 using real-time reverse transcription recombinase polymerase amplification BMC Veterinary Research Goose astrovirus genotypes 2 Real-time Reverse transcription recombinase polymerase amplification Point-of-care diagnosis |
| title | Rapid detection of goose astrovirus genotypes 2 using real-time reverse transcription recombinase polymerase amplification |
| title_full | Rapid detection of goose astrovirus genotypes 2 using real-time reverse transcription recombinase polymerase amplification |
| title_fullStr | Rapid detection of goose astrovirus genotypes 2 using real-time reverse transcription recombinase polymerase amplification |
| title_full_unstemmed | Rapid detection of goose astrovirus genotypes 2 using real-time reverse transcription recombinase polymerase amplification |
| title_short | Rapid detection of goose astrovirus genotypes 2 using real-time reverse transcription recombinase polymerase amplification |
| title_sort | rapid detection of goose astrovirus genotypes 2 using real time reverse transcription recombinase polymerase amplification |
| topic | Goose astrovirus genotypes 2 Real-time Reverse transcription recombinase polymerase amplification Point-of-care diagnosis |
| url | https://doi.org/10.1186/s12917-023-03790-2 |
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