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| Auteurs principaux: | , , , , , , , , , , |
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| Format: | Artículo científico |
| Langue: | en |
| Publié: |
Biology
2025
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| Accès en ligne: | https://pubmed.ncbi.nlm.nih.gov/40282195/ |
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| _version_ | 1868266211773513730 |
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| author | Chen, Minqi Zhou, Yongcan Wang, Shifeng Luo, Jian Guo, Weiliang Deng, Hengwei Zheng, Pei Zhong, Zhihong Su, Baofeng Zhang, Dongdong Ye, Zhi |
| author_facet | Chen, Minqi Zhou, Yongcan Wang, Shifeng Luo, Jian Guo, Weiliang Deng, Hengwei Zheng, Pei Zhong, Zhihong Su, Baofeng Zhang, Dongdong Ye, Zhi Chen, Minqi Zhou, Yongcan Wang, Shifeng Luo, Jian Guo, Weiliang Deng, Hengwei Zheng, Pei Zhong, Zhihong Su, Baofeng Zhang, Dongdong Ye, Zhi |
| collection | PubMed - marine biology |
| contents | Development of a Real-Time Enzymatic Recombinase Amplification Assay (RT-ERA) and an ERA Combined with a Lateral Flow Dipstick (LFD) Assay (ERA-LFD) for Enteric Microsporidian () in Grouper Fishes. Chen, Minqi Zhou, Yongcan Wang, Shifeng Luo, Jian Guo, Weiliang Deng, Hengwei Zheng, Pei Zhong, Zhihong Su, Baofeng Zhang, Dongdong Ye, Zhi poses a severe threat to grouper aquaculture due to the absence of effective prevention and treatment strategies. To address this challenge, we developed and validated two isothermal diagnostic tools, the real-time enzymatic recombinase amplification (RT-ERA) assay and the enzymatic recombinase amplification combined with a lateral flow dipstick (ERA-LFD) assay, targeting the 18S rDNA gene of the parasite. These assays operate under isothermal conditions at ≤40 °C and offer rapid detection, with RT-ERA yielding results in 14~20 min and ERA-LFD in approximately 10 min. The RT-ERA assay demonstrated a strong linear relationship between plasmid copy numbers and cycle threshold (Ct) values (y = -2.1226x + 19.562, R = 0.9915), enabling accurate quantification. Both methods displayed a detection limit of 2 × 10 copies/μL and no cross-reactivity with other aquaculture pathogens. Validation using grouper tissue and water samples from Hainan, China, demonstrated 100% concordance rates with basic ERA and outperformed compared to conventional PCR. These assays provide sensitive, specific, and rapid detection tools for effective monitoring and pathogen load assessment of , with broad applicability to pathogen detection in aquaculture systems. |
| format | Artículo científico |
| id | pubmed_40282195 |
| institution | PubMed |
| language | en |
| publishDate | 2025 |
| publisher | Biology |
| record_format | pubmed |
| spellingShingle | Development of a Real-Time Enzymatic Recombinase Amplification Assay (RT-ERA) and an ERA Combined with a Lateral Flow Dipstick (LFD) Assay (ERA-LFD) for Enteric Microsporidian () in Grouper Fishes. Chen, Minqi Zhou, Yongcan Wang, Shifeng Luo, Jian Guo, Weiliang Deng, Hengwei Zheng, Pei Zhong, Zhihong Su, Baofeng Zhang, Dongdong Ye, Zhi Development of a Real-Time Enzymatic Recombinase Amplification Assay (RT-ERA) and an ERA Combined with a Lateral Flow Dipstick (LFD) Assay (ERA-LFD) for Enteric Microsporidian () in Grouper Fishes. Chen, Minqi Zhou, Yongcan Wang, Shifeng Luo, Jian Guo, Weiliang Deng, Hengwei Zheng, Pei Zhong, Zhihong Su, Baofeng Zhang, Dongdong Ye, Zhi poses a severe threat to grouper aquaculture due to the absence of effective prevention and treatment strategies. To address this challenge, we developed and validated two isothermal diagnostic tools, the real-time enzymatic recombinase amplification (RT-ERA) assay and the enzymatic recombinase amplification combined with a lateral flow dipstick (ERA-LFD) assay, targeting the 18S rDNA gene of the parasite. These assays operate under isothermal conditions at ≤40 °C and offer rapid detection, with RT-ERA yielding results in 14~20 min and ERA-LFD in approximately 10 min. The RT-ERA assay demonstrated a strong linear relationship between plasmid copy numbers and cycle threshold (Ct) values (y = -2.1226x + 19.562, R = 0.9915), enabling accurate quantification. Both methods displayed a detection limit of 2 × 10 copies/μL and no cross-reactivity with other aquaculture pathogens. Validation using grouper tissue and water samples from Hainan, China, demonstrated 100% concordance rates with basic ERA and outperformed compared to conventional PCR. These assays provide sensitive, specific, and rapid detection tools for effective monitoring and pathogen load assessment of , with broad applicability to pathogen detection in aquaculture systems. |
| title | Development of a Real-Time Enzymatic Recombinase Amplification Assay (RT-ERA) and an ERA Combined with a Lateral Flow Dipstick (LFD) Assay (ERA-LFD) for Enteric Microsporidian () in Grouper Fishes. |
| url | https://pubmed.ncbi.nlm.nih.gov/40282195/ |