ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Infectious Agents and Disease
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1607974
This article is part of the Research TopicRapid and Efficient Analytical Technologies for Pathogen DetectionView all 5 articles
Recombinase polymerase amplification combined with CRISPR/Cas12a technology for rapid on-site detection of duck adenovirus 3
Provisionally accepted
Qizhang Liang1*
Wei Chen1Yuhai Bi2
Weiwei Wang1Rong-Chang Liu1Qiu-Ling Fu1Guang-Hua Fu1Long-Fei Cheng1Hong-Mei Chen1Nan-Song Jiang1Ting Zhu3Yu Huang1- 1Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou, China
- 2Institute of Microbiology, Chinese Academy of Sciences (CAS), Beijing, Beijing Municipality, China
- 3College of Animal Science, Fujian agriculture and Forestry University, Fuzhou, Fujian Province, China
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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°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 minutes.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.
Keywords: DAdV-3, RPA, LFS, CRISPR/Cas12a, on-site detection
Received: 10 Apr 2025; Accepted: 21 Jul 2025.
Copyright: © 2025 Liang, Chen, Bi, Wang, Liu, Fu, Fu, Cheng, Chen, Jiang, Zhu and Huang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Qizhang Liang, Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou, China
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