Development and application of a reverse-transcription recombinase-aided amplification assay for subgroup J Avian leukosis virus
ABSTRACT: Subgroup J Avian leukosis virus (ALV-J) is an important pathogen of poultry tumor diseases. Since its discovery, it has caused significant economic losses to the poultry industry. Thus, the rapid detection of molecular level with strong specificity is particularly important whether poultry...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-04-01
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| Series: | Poultry Science |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0032579122000487 |
| _version_ | 1819022394484850688 |
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| author | X.H. Wu Z.Q. Yao Q.Q. Zhao S. Chen Z.Z. Hu Z. Xie L.Y. Chen J. Ji F. Chen X.H. Zhang Q.M. Xie |
| author_facet | X.H. Wu Z.Q. Yao Q.Q. Zhao S. Chen Z.Z. Hu Z. Xie L.Y. Chen J. Ji F. Chen X.H. Zhang Q.M. Xie |
| author_sort | X.H. Wu |
| collection | DOAJ |
| description | ABSTRACT: Subgroup J Avian leukosis virus (ALV-J) is an important pathogen of poultry tumor diseases. Since its discovery, it has caused significant economic losses to the poultry industry. Thus, the rapid detection of molecular level with strong specificity is particularly important whether poultry are infected with ALV-J. In this study, we designed primers and probe for real-time fluorescent reverse-transcription recombinase-aided amplification assay (RT-RAA) based on the ALV-J gp85 sequence. We had established a real-time fluorescent RT-RAA method and confirmed this system by verifying the specificity and sensitivity of the primers and probe. In addition, repeatability tests and clinical sample regression tests were used for preliminary evaluation of this detection method. The sensitivity of established method was about 101 copies/μL, and the repeatability of the CV of the CT value is 4%, indicating repeatability is good. Moreover, there was no cross-reactivity with NDV, IBV, IBDV, H9N2, MDV, and REV, and other avian leukosis virus subgroups, such as subgroups A, B, C, D, K and E. Importantly, the real-time fluorescent RT-RAA completed the test within 30 min at a constant temperature of 41°C. Forty-two clinical samples with known background were tested, and the test results were coincided with 100%. Overall, these results suggested that the real-time fluorescent RT-RAA developed in this study had strong specificity, high sensitivity, and good feasibility. The method is simple, easy, and portable, that is suitable for clinical and laboratory diagnosis, and provides technical support for the prevention and control of ALV-J. |
| first_indexed | 2024-12-21T04:22:18Z |
| format | Article |
| id | doaj.art-9feb0aeea16b419daaefbb538fd0f1bd |
| institution | Directory Open Access Journal |
| issn | 0032-5791 |
| language | English |
| last_indexed | 2024-12-21T04:22:18Z |
| publishDate | 2022-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Poultry Science |
| spelling | doaj.art-9feb0aeea16b419daaefbb538fd0f1bd2022-12-21T19:16:08ZengElsevierPoultry Science0032-57912022-04-011014101743Development and application of a reverse-transcription recombinase-aided amplification assay for subgroup J Avian leukosis virusX.H. Wu0Z.Q. Yao1Q.Q. Zhao2S. Chen3Z.Z. Hu4Z. Xie5L.Y. Chen6J. Ji7F. Chen8X.H. Zhang9Q.M. Xie10Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. ChinaHeyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. ChinaHeyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. ChinaHeyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. ChinaHeyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. ChinaHeyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. ChinaHeyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. ChinaHenan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, Nanyang Normal University, Nanyang 473061, P. R. ChinaHeyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. China; Key Laboratory of Animal Health Aquaculture and Environmental Control, Guangzhou, Guangdong 510642, P. R. ChinaHeyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. China; Key Laboratory of Animal Health Aquaculture and Environmental Control, Guangzhou, Guangdong 510642, P. R. ChinaHeyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, P. R. China; Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou 510642, P. R. China; South China Collaborative Innovation Center for Poultry Disease Control and Product Safety, Guangzhou 510642, P. R. China; Key Laboratory of Animal Health Aquaculture and Environmental Control, Guangzhou, Guangdong 510642, P. R. China; Corresponding author:ABSTRACT: Subgroup J Avian leukosis virus (ALV-J) is an important pathogen of poultry tumor diseases. Since its discovery, it has caused significant economic losses to the poultry industry. Thus, the rapid detection of molecular level with strong specificity is particularly important whether poultry are infected with ALV-J. In this study, we designed primers and probe for real-time fluorescent reverse-transcription recombinase-aided amplification assay (RT-RAA) based on the ALV-J gp85 sequence. We had established a real-time fluorescent RT-RAA method and confirmed this system by verifying the specificity and sensitivity of the primers and probe. In addition, repeatability tests and clinical sample regression tests were used for preliminary evaluation of this detection method. The sensitivity of established method was about 101 copies/μL, and the repeatability of the CV of the CT value is 4%, indicating repeatability is good. Moreover, there was no cross-reactivity with NDV, IBV, IBDV, H9N2, MDV, and REV, and other avian leukosis virus subgroups, such as subgroups A, B, C, D, K and E. Importantly, the real-time fluorescent RT-RAA completed the test within 30 min at a constant temperature of 41°C. Forty-two clinical samples with known background were tested, and the test results were coincided with 100%. Overall, these results suggested that the real-time fluorescent RT-RAA developed in this study had strong specificity, high sensitivity, and good feasibility. The method is simple, easy, and portable, that is suitable for clinical and laboratory diagnosis, and provides technical support for the prevention and control of ALV-J.http://www.sciencedirect.com/science/article/pii/S0032579122000487subgroup J Avian leukosis virusreverse-transcription recombinase-aided amplification assaygp85constant temperature detection |
| spellingShingle | X.H. Wu Z.Q. Yao Q.Q. Zhao S. Chen Z.Z. Hu Z. Xie L.Y. Chen J. Ji F. Chen X.H. Zhang Q.M. Xie Development and application of a reverse-transcription recombinase-aided amplification assay for subgroup J Avian leukosis virus Poultry Science subgroup J Avian leukosis virus reverse-transcription recombinase-aided amplification assay gp85 constant temperature detection |
| title | Development and application of a reverse-transcription recombinase-aided amplification assay for subgroup J Avian leukosis virus |
| title_full | Development and application of a reverse-transcription recombinase-aided amplification assay for subgroup J Avian leukosis virus |
| title_fullStr | Development and application of a reverse-transcription recombinase-aided amplification assay for subgroup J Avian leukosis virus |
| title_full_unstemmed | Development and application of a reverse-transcription recombinase-aided amplification assay for subgroup J Avian leukosis virus |
| title_short | Development and application of a reverse-transcription recombinase-aided amplification assay for subgroup J Avian leukosis virus |
| title_sort | development and application of a reverse transcription recombinase aided amplification assay for subgroup j avian leukosis virus |
| topic | subgroup J Avian leukosis virus reverse-transcription recombinase-aided amplification assay gp85 constant temperature detection |
| url | http://www.sciencedirect.com/science/article/pii/S0032579122000487 |
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