Rapid and visual detection of novel astroviruses causing fatal gout in goslings using one-step reverse transcription loop-mediated isothermal amplification
To visually and rapidly detect a novel goose astrovirus (N-GoAstV) causing fatal gout in goslings, an isothermal detection method based on one-step reverse transcription loop-mediated isothermal amplification (one-step RT-LAMP) was established. The one-step RT-LAMP assay for N-GoAstV detection, usin...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-09-01
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| Series: | Poultry Science |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S003257912030328X |
| _version_ | 1818156935504986112 |
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| author | Jun Ji Qinxi Chen Chaoge Sui Wen Hu Zhengli Yu Zhibin Zhang Xinhao Mu Xin Xu Lunguang Yao Yunchao Kan Qingmei Xie |
| author_facet | Jun Ji Qinxi Chen Chaoge Sui Wen Hu Zhengli Yu Zhibin Zhang Xinhao Mu Xin Xu Lunguang Yao Yunchao Kan Qingmei Xie |
| author_sort | Jun Ji |
| collection | DOAJ |
| description | To visually and rapidly detect a novel goose astrovirus (N-GoAstV) causing fatal gout in goslings, an isothermal detection method based on one-step reverse transcription loop-mediated isothermal amplification (one-step RT-LAMP) was established. The one-step RT-LAMP assay for N-GoAstV detection, using Bst 3.0 DNA polymerase with strong reverse transcription activity and primer sets targeting the opening reading frame 1b (ORF1b) of N-GoAstV, could be completed in 30 min using a water bath at 61°C; the detection results could be visually observed by adding a pH-sensitive dye containing phenol red and cresol red. The detection limit of the one-step RT-LAMP assay was 57.8 copies, which was similar to that of reverse transcription–quantitative polymerase chain reaction. The assay specifically detected N-GoAstV without any cross-reaction with other reference viruses, and this was further confirmed using enzyme digestion. These results indicated that the newly established RT-LAMP assay could accomplish reverse transcription, amplification, and visual result determination in one step, and the results obtained via this rapid and cost-effective method could be used to support disease control on farms in terms of N-GoAstV infection. |
| first_indexed | 2024-12-11T15:06:12Z |
| format | Article |
| id | doaj.art-4158f644657c4556b1cc0e969d5bc513 |
| institution | Directory Open Access Journal |
| issn | 0032-5791 |
| language | English |
| last_indexed | 2024-12-11T15:06:12Z |
| publishDate | 2020-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Poultry Science |
| spelling | doaj.art-4158f644657c4556b1cc0e969d5bc5132022-12-22T01:00:54ZengElsevierPoultry Science0032-57912020-09-0199942594264Rapid and visual detection of novel astroviruses causing fatal gout in goslings using one-step reverse transcription loop-mediated isothermal amplificationJun Ji0Qinxi Chen1Chaoge Sui2Wen Hu3Zhengli Yu4Zhibin Zhang5Xinhao Mu6Xin Xu7Lunguang Yao8Yunchao Kan9Qingmei Xie10Henan Provincial Engineering Laboratory of Insects Bio-Reactor, Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, Nanyang Normal University, Nanyang 473061, PR China; Corresponding authors:Henan Provincial Engineering Laboratory of Insects Bio-Reactor, Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, Nanyang Normal University, Nanyang 473061, PR ChinaHenan Provincial Engineering Laboratory of Insects Bio-Reactor, Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, Nanyang Normal University, Nanyang 473061, PR ChinaHenan Provincial Engineering Laboratory of Insects Bio-Reactor, Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, Nanyang Normal University, Nanyang 473061, PR ChinaHenan Provincial Engineering Laboratory of Insects Bio-Reactor, Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, Nanyang Normal University, Nanyang 473061, PR ChinaHenan Provincial Engineering Laboratory of Insects Bio-Reactor, Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, Nanyang Normal University, Nanyang 473061, PR ChinaHenan Provincial Engineering Laboratory of Insects Bio-Reactor, Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, Nanyang Normal University, Nanyang 473061, PR ChinaHenan Provincial Engineering Laboratory of Insects Bio-Reactor, Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, Nanyang Normal University, Nanyang 473061, PR China; Corresponding authors:Henan Provincial Engineering Laboratory of Insects Bio-Reactor, Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, Nanyang Normal University, Nanyang 473061, PR China; Corresponding authors:Henan Provincial Engineering Laboratory of Insects Bio-Reactor, Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, Nanyang Normal University, Nanyang 473061, PR ChinaCollege of Animal Science, South China Agricultural University, Guangzhou 510642, PR ChinaTo visually and rapidly detect a novel goose astrovirus (N-GoAstV) causing fatal gout in goslings, an isothermal detection method based on one-step reverse transcription loop-mediated isothermal amplification (one-step RT-LAMP) was established. The one-step RT-LAMP assay for N-GoAstV detection, using Bst 3.0 DNA polymerase with strong reverse transcription activity and primer sets targeting the opening reading frame 1b (ORF1b) of N-GoAstV, could be completed in 30 min using a water bath at 61°C; the detection results could be visually observed by adding a pH-sensitive dye containing phenol red and cresol red. The detection limit of the one-step RT-LAMP assay was 57.8 copies, which was similar to that of reverse transcription–quantitative polymerase chain reaction. The assay specifically detected N-GoAstV without any cross-reaction with other reference viruses, and this was further confirmed using enzyme digestion. These results indicated that the newly established RT-LAMP assay could accomplish reverse transcription, amplification, and visual result determination in one step, and the results obtained via this rapid and cost-effective method could be used to support disease control on farms in terms of N-GoAstV infection.http://www.sciencedirect.com/science/article/pii/S003257912030328XN-GoAstVrapidityvisible detectionone-step RT-LAMP |
| spellingShingle | Jun Ji Qinxi Chen Chaoge Sui Wen Hu Zhengli Yu Zhibin Zhang Xinhao Mu Xin Xu Lunguang Yao Yunchao Kan Qingmei Xie Rapid and visual detection of novel astroviruses causing fatal gout in goslings using one-step reverse transcription loop-mediated isothermal amplification Poultry Science N-GoAstV rapidity visible detection one-step RT-LAMP |
| title | Rapid and visual detection of novel astroviruses causing fatal gout in goslings using one-step reverse transcription loop-mediated isothermal amplification |
| title_full | Rapid and visual detection of novel astroviruses causing fatal gout in goslings using one-step reverse transcription loop-mediated isothermal amplification |
| title_fullStr | Rapid and visual detection of novel astroviruses causing fatal gout in goslings using one-step reverse transcription loop-mediated isothermal amplification |
| title_full_unstemmed | Rapid and visual detection of novel astroviruses causing fatal gout in goslings using one-step reverse transcription loop-mediated isothermal amplification |
| title_short | Rapid and visual detection of novel astroviruses causing fatal gout in goslings using one-step reverse transcription loop-mediated isothermal amplification |
| title_sort | rapid and visual detection of novel astroviruses causing fatal gout in goslings using one step reverse transcription loop mediated isothermal amplification |
| topic | N-GoAstV rapidity visible detection one-step RT-LAMP |
| url | http://www.sciencedirect.com/science/article/pii/S003257912030328X |
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