Rapid and visual detection of Mycoplasma synoviae by recombinase-aided amplification assay combined with a lateral flow dipstick
Mycoplasma synoviae (MS) is an important avian pathogen that has brought substantial economic losses to the global poultry industry. Fast and accurate diagnosis is one of the critical factors for the control of MS infection. This study established a simple, rapid and visual detection method for MS u...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-07-01
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| Series: | Poultry Science |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0032579122001675 |
| _version_ | 1818231712501465088 |
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| author | Wenlong Xia Ke Chen Wensong Liu Yan Yin Qian Yao Yu Ban Yiwen Pu Xingmin Zhan Hongchun Bian Shupei Yu Kunpeng Han Ling Yang Huanli Wang Zhongjun Fan |
| author_facet | Wenlong Xia Ke Chen Wensong Liu Yan Yin Qian Yao Yu Ban Yiwen Pu Xingmin Zhan Hongchun Bian Shupei Yu Kunpeng Han Ling Yang Huanli Wang Zhongjun Fan |
| author_sort | Wenlong Xia |
| collection | DOAJ |
| description | Mycoplasma synoviae (MS) is an important avian pathogen that has brought substantial economic losses to the global poultry industry. Fast and accurate diagnosis is one of the critical factors for the control of MS infection. This study established a simple, rapid and visual detection method for MS using a recombinase-aided amplification (RAA) combined with a lateral flow dipstick (LFD). The reaction temperature and time of the RAA-LFD assay were optimized after selecting the primers and probe, and the specificity and sensitivity rates were analyzed. The results showed that RAA could amplify the target gene in 20 min at a constant temperature of 38°C, and the amplification products could be visualized by LFD within 5 min. There was no cross-reaction with Mycoplasma gallisepticum (MG), Pasteurella multocida (P. multocida), Escherichia coli (E. coli), Newcastle disease virus (NDV), infectious bursal disease virus (IBDV), infectious bronchitis virus (IBV), and avian reovirus (ARV). Furthermore, the RAA-LFD assay exhibited high sensitivity with a detection limit of 10 copies/μL. A total of 128 clinical samples with suspected infection of MS were tested by RAA-LFD, PCR, and real-time fluorescence quantitative PCR (RFQ-PCR). The coincidence rate of the detection results was 95.3% between RAA-LFD and PCR, and 98.4% between RAA-LFD and RFQ-PCR. These results suggested that the RAA-LFD method established in the present study was easy to use and was associated with strong specificity and high sensitivity. This method was very suitable for the rapid detection of MS in clinical practice. |
| first_indexed | 2024-12-12T10:54:45Z |
| format | Article |
| id | doaj.art-197d995a0ecc44ef8044cee5bb3d64cf |
| institution | Directory Open Access Journal |
| issn | 0032-5791 |
| language | English |
| last_indexed | 2024-12-12T10:54:45Z |
| publishDate | 2022-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Poultry Science |
| spelling | doaj.art-197d995a0ecc44ef8044cee5bb3d64cf2022-12-22T00:26:41ZengElsevierPoultry Science0032-57912022-07-011017101860Rapid and visual detection of Mycoplasma synoviae by recombinase-aided amplification assay combined with a lateral flow dipstickWenlong Xia0Ke Chen1Wensong Liu2Yan Yin3Qian Yao4Yu Ban5Yiwen Pu6Xingmin Zhan7Hongchun Bian8Shupei Yu9Kunpeng Han10Ling Yang11Huanli Wang12Zhongjun Fan13Yancheng Engineering Research Center of Animal Biologics, College of Marine and Biological Engineering, Yancheng Teachers University, Yancheng 224002, ChinaYancheng Engineering Research Center of Animal Biologics, College of Marine and Biological Engineering, Yancheng Teachers University, Yancheng 224002, ChinaYancheng Engineering Research Center of Animal Biologics, College of Marine and Biological Engineering, Yancheng Teachers University, Yancheng 224002, ChinaYancheng Engineering Research Center of Animal Biologics, College of Marine and Biological Engineering, Yancheng Teachers University, Yancheng 224002, ChinaYancheng Engineering Research Center of Animal Biologics, College of Marine and Biological Engineering, Yancheng Teachers University, Yancheng 224002, ChinaYancheng Engineering Research Center of Animal Biologics, College of Marine and Biological Engineering, Yancheng Teachers University, Yancheng 224002, ChinaYancheng Engineering Research Center of Animal Biologics, College of Marine and Biological Engineering, Yancheng Teachers University, Yancheng 224002, ChinaYancheng Engineering Research Center of Animal Biologics, College of Marine and Biological Engineering, Yancheng Teachers University, Yancheng 224002, ChinaYancheng Animal Husbandry and Veterinary Station, Yancheng 224001, ChinaYancheng Animal Husbandry and Veterinary Station, Yancheng 224001, ChinaYancheng Animal Husbandry and Veterinary Station, Yancheng 224001, ChinaYancheng Animal Husbandry and Veterinary Station, Yancheng 224001, ChinaYancheng Engineering Research Center of Animal Biologics, College of Marine and Biological Engineering, Yancheng Teachers University, Yancheng 224002, ChinaYancheng Engineering Research Center of Animal Biologics, College of Marine and Biological Engineering, Yancheng Teachers University, Yancheng 224002, China; Corresponding author:Mycoplasma synoviae (MS) is an important avian pathogen that has brought substantial economic losses to the global poultry industry. Fast and accurate diagnosis is one of the critical factors for the control of MS infection. This study established a simple, rapid and visual detection method for MS using a recombinase-aided amplification (RAA) combined with a lateral flow dipstick (LFD). The reaction temperature and time of the RAA-LFD assay were optimized after selecting the primers and probe, and the specificity and sensitivity rates were analyzed. The results showed that RAA could amplify the target gene in 20 min at a constant temperature of 38°C, and the amplification products could be visualized by LFD within 5 min. There was no cross-reaction with Mycoplasma gallisepticum (MG), Pasteurella multocida (P. multocida), Escherichia coli (E. coli), Newcastle disease virus (NDV), infectious bursal disease virus (IBDV), infectious bronchitis virus (IBV), and avian reovirus (ARV). Furthermore, the RAA-LFD assay exhibited high sensitivity with a detection limit of 10 copies/μL. A total of 128 clinical samples with suspected infection of MS were tested by RAA-LFD, PCR, and real-time fluorescence quantitative PCR (RFQ-PCR). The coincidence rate of the detection results was 95.3% between RAA-LFD and PCR, and 98.4% between RAA-LFD and RFQ-PCR. These results suggested that the RAA-LFD method established in the present study was easy to use and was associated with strong specificity and high sensitivity. This method was very suitable for the rapid detection of MS in clinical practice.http://www.sciencedirect.com/science/article/pii/S0032579122001675Mycoplasma synoviaerecombinase-aided amplificationlateral flow dipstickPCRreal-time fluorescence quantitative PCR |
| spellingShingle | Wenlong Xia Ke Chen Wensong Liu Yan Yin Qian Yao Yu Ban Yiwen Pu Xingmin Zhan Hongchun Bian Shupei Yu Kunpeng Han Ling Yang Huanli Wang Zhongjun Fan Rapid and visual detection of Mycoplasma synoviae by recombinase-aided amplification assay combined with a lateral flow dipstick Poultry Science Mycoplasma synoviae recombinase-aided amplification lateral flow dipstick PCR real-time fluorescence quantitative PCR |
| title | Rapid and visual detection of Mycoplasma synoviae by recombinase-aided amplification assay combined with a lateral flow dipstick |
| title_full | Rapid and visual detection of Mycoplasma synoviae by recombinase-aided amplification assay combined with a lateral flow dipstick |
| title_fullStr | Rapid and visual detection of Mycoplasma synoviae by recombinase-aided amplification assay combined with a lateral flow dipstick |
| title_full_unstemmed | Rapid and visual detection of Mycoplasma synoviae by recombinase-aided amplification assay combined with a lateral flow dipstick |
| title_short | Rapid and visual detection of Mycoplasma synoviae by recombinase-aided amplification assay combined with a lateral flow dipstick |
| title_sort | rapid and visual detection of mycoplasma synoviae by recombinase aided amplification assay combined with a lateral flow dipstick |
| topic | Mycoplasma synoviae recombinase-aided amplification lateral flow dipstick PCR real-time fluorescence quantitative PCR |
| url | http://www.sciencedirect.com/science/article/pii/S0032579122001675 |
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