ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Virology
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1638182
Development of a rapid recombinase polymerase amplification-lateral flow dipstick assay for sensitive detection of duck adenovirus type-3
Provisionally accepted
Jiayu Sun1
Kewei Liu1Chao Liu1Zhenyu Wang1Jiaxi Gao1Ruya Zhao1Long Yuan1Yan Shen2
Jinchun Li1
Fangfang Chen1*- 1Anhui Agricultural University, Hefei, China
- 2Anhui Animal Disease Prevention and Control Center, Hefei, China
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Duck adenovirus type-3 (DAdV-3) infections have severe effects on duck health, and rapid detection methods are crucial to reduce the morbidity and mortality associated with this pathogen in clinical practice. In this study, we established, optimized, and validated a novel recombinase polymerase amplification (RPA)-lateral flow dipstick (LFD) assay for the detection of DAdV-3. Next, we established a clinical infection model based on the pathogenicity of the DAdV-3 strain and tested the effectiveness of the RPA-LFD assay in controlling an outbreak of DAdV-3 infection. The findings indicated that the RPA-LFD assay could be performed within 30 minutes at 42 ℃. Specificity tests indicated no cross-reactivity with other viruses. The detection limit of the assay was 1 × 101 copies/µL. We evaluated 65 clinical samples using RPA-LFD and quantitative polymerase chain reaction (qPCR), and both methods showed a positivity rate of 33.8% and a coincidence rate of 100%. The kappa (κ) value of the RPA-LFD and qPCR assays was 1 (p < 0.001). The application of this assay in experimentally infected ducklings reduced the mortality rate from 20% to 8%. Thus, the RPA-LFD assay established in this study demonstrated high specificity, sensitivity, rapidity, and efficacy, indicating its potential for rapid detection of DAdV-3 in clinical settings.
Keywords: DAdV-3, RPA-LFD, detection, clinical diagnosis, specificity
Received: 30 May 2025; Accepted: 31 Jul 2025.
Copyright: © 2025 Sun, Liu, Liu, Wang, Gao, Zhao, Yuan, Shen, Li and Chen. 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: Fangfang Chen, Anhui Agricultural University, Hefei, China
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