Recombinase polymerase amplification combined with CRISPR/Cas12a technology for rapid on-site detection of duck adenovirus 3
文献类型： 外文期刊
作者： Liang, Qi-Zhang 1 ; Chen, Wei 1 ; Bi, Yuhai 3 ; Wang, Weiwei 1 ; Liu, Rong-Chang 1 ; Fu, Qiu-Ling 1 ; Fu, Guang-Hua 1 ; Cheng, Long-Fei 1 ; Jiang, Nan-Song 1 ; Zhu, Ting 2 ; Chen, Hong-Mei 1 ; Huang, Yu 1 ;
作者机构： 1.Fujian Acad Agr Sci, Inst Anim Husb & Vet Med, Fujian Prov Key Lab Avian Dis Control & Prevent, Fuzhou, Peoples R China
2.Fujian Agr & Forestry Univ, Coll Anim Sci, Fuzhou, Peoples R China
3.Chinese Acad Sci, Inst Microbiol, CAS Key Lab Pathogen Microbiol & Immunol, Beijing, Peoples R China
关键词： DAdV-3; RPA; LFS; CRISPR/Cas12a; on-site detection
期刊名称：FRONTIERS IN MICROBIOLOGY （ 影响因子：4.5； 五年影响因子：5.2 ）
ISSN：
年卷期： 2025 年 16 卷
页码：
收录情况： SCI
摘要： Duck adenovirus 3 (DAdV-3) causes liver damage and bleeding, with morbidity rates ranging from 40 to 55% and mortality rates between 35 and 43%. Co-infection with other pathogens complicates disease control, significantly impacting the duck breeding industry. Currently, there have been no effective vaccines or treatments for DAdV-3. Therefore, rapid, specific, and sensitive detection methods are crucial for preventing and controlling this virus. Our study developed a lateral flow strip (LFS) detection method using recombinase polymerase amplification (RPA) and CRISPR/Cas12a. The RPA-CRISPR/Cas12a-LFS method, performed at 37 degrees C, allowed for result visualization without sophisticated equipment. It targeted the DAdV-3 Fiber-2 gene and achieved a detection limit of 3.0 gene copies. Additionally, this method demonstrated high specificity, with no cross-reactivity to eight other avian viruses. The reaction time of RPA-CRISPR/Cas12a-LFS is only 45 min. Analysis of 95 waterfowl samples showed 98.95% consistency and agreement with quantitative polymerase chain reaction using the Fiber-2 RPA-CRISPR/Cas12a-LFS method. These findings highlighted the potential of this user-friendly, rapid, sensitive, and accurate detection method for on-site DAdV-3 detection.
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