Establishment and Application Prospect of Reverse Transcriptase Recombinase-Aided Amplification Assay for Subgroup C Avian Metapneumovirus
Among broilers, the main pathogen that leads to swollen head syndrome (SHS) is the subgroup C avian metapneumovirus (aMPV-C). The aMPV-C infection can lead to an upsurge in the rate of soft-shell eggs, resulting in reduced egg production and seriously affecting the economy of the livestock industry....
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-03-01
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| Series: | Veterinary Sciences |
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| Online Access: | https://www.mdpi.com/2306-7381/11/3/122 |
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| author | Yuhang Bai Xiuhong Wu Jiajia Liu Zhanxin Wang Mengyue Dong Tong Li Zhenkai Dai Hongxin Li Qingmei Xie Xinheng Zhang |
| author_facet | Yuhang Bai Xiuhong Wu Jiajia Liu Zhanxin Wang Mengyue Dong Tong Li Zhenkai Dai Hongxin Li Qingmei Xie Xinheng Zhang |
| author_sort | Yuhang Bai |
| collection | DOAJ |
| description | Among broilers, the main pathogen that leads to swollen head syndrome (SHS) is the subgroup C avian metapneumovirus (aMPV-C). The aMPV-C infection can lead to an upsurge in the rate of soft-shell eggs, resulting in reduced egg production and seriously affecting the economy of the livestock industry. Therefore, a rapid method for aMPV-C detection needs to be invented. According to the N gene of aMPV-C, we designed the specific probe and primer and created a reverse transcription recombinase-aided amplification assay (RT-RAA) for the detection of aMPV-C. aMPV-C could be detected quickly and specifically by this method at 41 °C for 30 min. The sensitivity assay inferred that the minimum detection threshold of RT-RAA was 3.38 × 10<sup>1</sup> copies/μL. A specificity assay showed that the RT-RAA method did not cross-react with other subgroups (aMPV-A, aMPV-B, aMPV-D) or other viruses (H9N2, NDV, IBV, IBDV). Forty samples of known clinical background were tested by RT-RAA and RT-qPCR. The two approaches had a 100% correlation rate. In conclusion, this research successfully created an RT-RAA assay for aMPV-C. |
| first_indexed | 2024-04-24T17:46:12Z |
| format | Article |
| id | doaj.art-78e394a97e1d4cba8ca96ad714562bbc |
| institution | Directory Open Access Journal |
| issn | 2306-7381 |
| language | English |
| last_indexed | 2024-04-24T17:46:12Z |
| publishDate | 2024-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Veterinary Sciences |
| spelling | doaj.art-78e394a97e1d4cba8ca96ad714562bbc2024-03-27T14:07:23ZengMDPI AGVeterinary Sciences2306-73812024-03-0111312210.3390/vetsci11030122Establishment and Application Prospect of Reverse Transcriptase Recombinase-Aided Amplification Assay for Subgroup C Avian MetapneumovirusYuhang Bai0Xiuhong Wu1Jiajia Liu2Zhanxin Wang3Mengyue Dong4Tong Li5Zhenkai Dai6Hongxin Li7Qingmei Xie8Xinheng Zhang9State Key Laboratory of Swine and Poultry Breeding Industry & Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, ChinaState Key Laboratory of Swine and Poultry Breeding Industry & Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, ChinaGuangdong Wen’s Foodstuffs Group Co., Ltd., Yunfu 527439, ChinaGuangdong Wen’s Foodstuffs Group Co., Ltd., Yunfu 527439, ChinaState Key Laboratory of Swine and Poultry Breeding Industry & Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, ChinaState Key Laboratory of Swine and Poultry Breeding Industry & Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, ChinaState Key Laboratory of Swine and Poultry Breeding Industry & Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, ChinaState Key Laboratory of Swine and Poultry Breeding Industry & Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, ChinaState Key Laboratory of Swine and Poultry Breeding Industry & Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, ChinaState Key Laboratory of Swine and Poultry Breeding Industry & Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, ChinaAmong broilers, the main pathogen that leads to swollen head syndrome (SHS) is the subgroup C avian metapneumovirus (aMPV-C). The aMPV-C infection can lead to an upsurge in the rate of soft-shell eggs, resulting in reduced egg production and seriously affecting the economy of the livestock industry. Therefore, a rapid method for aMPV-C detection needs to be invented. According to the N gene of aMPV-C, we designed the specific probe and primer and created a reverse transcription recombinase-aided amplification assay (RT-RAA) for the detection of aMPV-C. aMPV-C could be detected quickly and specifically by this method at 41 °C for 30 min. The sensitivity assay inferred that the minimum detection threshold of RT-RAA was 3.38 × 10<sup>1</sup> copies/μL. A specificity assay showed that the RT-RAA method did not cross-react with other subgroups (aMPV-A, aMPV-B, aMPV-D) or other viruses (H9N2, NDV, IBV, IBDV). Forty samples of known clinical background were tested by RT-RAA and RT-qPCR. The two approaches had a 100% correlation rate. In conclusion, this research successfully created an RT-RAA assay for aMPV-C.https://www.mdpi.com/2306-7381/11/3/122subgroup C avian metapneumovirusreverse transcriptase recombinase-aided amplification assayN geneconstant temperature detection |
| spellingShingle | Yuhang Bai Xiuhong Wu Jiajia Liu Zhanxin Wang Mengyue Dong Tong Li Zhenkai Dai Hongxin Li Qingmei Xie Xinheng Zhang Establishment and Application Prospect of Reverse Transcriptase Recombinase-Aided Amplification Assay for Subgroup C Avian Metapneumovirus Veterinary Sciences subgroup C avian metapneumovirus reverse transcriptase recombinase-aided amplification assay N gene constant temperature detection |
| title | Establishment and Application Prospect of Reverse Transcriptase Recombinase-Aided Amplification Assay for Subgroup C Avian Metapneumovirus |
| title_full | Establishment and Application Prospect of Reverse Transcriptase Recombinase-Aided Amplification Assay for Subgroup C Avian Metapneumovirus |
| title_fullStr | Establishment and Application Prospect of Reverse Transcriptase Recombinase-Aided Amplification Assay for Subgroup C Avian Metapneumovirus |
| title_full_unstemmed | Establishment and Application Prospect of Reverse Transcriptase Recombinase-Aided Amplification Assay for Subgroup C Avian Metapneumovirus |
| title_short | Establishment and Application Prospect of Reverse Transcriptase Recombinase-Aided Amplification Assay for Subgroup C Avian Metapneumovirus |
| title_sort | establishment and application prospect of reverse transcriptase recombinase aided amplification assay for subgroup c avian metapneumovirus |
| topic | subgroup C avian metapneumovirus reverse transcriptase recombinase-aided amplification assay N gene constant temperature detection |
| url | https://www.mdpi.com/2306-7381/11/3/122 |
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