ORIGINAL RESEARCH article
Front. Vet. Sci.
Sec. Veterinary Clinical, Anatomical, and Comparative Pathology
Development of a Colloidal Gold Immunochromatographic Strip for the Rapid Detection of Fowl Adenovirus Serotype 4 Using Prepared Penton Protein-Specific Monoclonal Antibodies
Provisionally accepted
Sisi Luo1,2
Bingyi Yang2
Jiaoling Huang2
You Wei2
Zhixun Xie2*
Xiaofeng Li2Aiqiong Wu2Zhihua Ruan2
Sheng Wang2
Yanfang Zhang2
Meng Li2Liji Xie2Ming Yan2
Weiwei Wang3Ping Wei3*- 1Institute for Poultry Science and Health，College of Animal Science and Technology, Guangxi University, Nanning, China
- 2Guangxi Veterinary Research Institute, Nanning, China
- 3Guangxi University, Nanning, China
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Fowl adenovirus serotype 4 (FAdV-4) is the primary pathogen responsible for hydropericardium-hepatitis syndrome (HHS) and is associated with high mortality rates (20–80%) in 3–6-week-old chickens. This study aimed to develop a rapid and specific method for detecting FAdV-4. Two monoclonal antibodies (mAbs 6B3 and 8G11) against the FAdV-4 penton protein were successfully generated using hybridoma technology, both of which exhibited high titers (1:100,000) and strong serotype specificity. Specificity analysis confirmed that these mAbs recognized 12 FAdV-4 isolates from diverse origins without cross-reactivity to other FAdV serotypes or common avian pathogens. On this basis, a colloidal gold immunochromatographic assay was developed; systematic optimization of its key parameters yielded an optimal labeling pH of 8.3, an antibody labeling concentration of 7.2 μg/mL, and optimal coating concentrations for the test line (T-line) and control line (C-line) of 2.5 mg/mL and 1.5 mg/mL, respectively. Performance evaluation of the method demonstrated that it achieved a detection sensitivity of 7.81×104 TCID₅₀/0.1 mL with a detection time of 15 minutes. Clinical sample validation revealed a positive concordance rate of 91.6%, a negative concordance rate of 100%, and an overall concordance rate of 98.6% relative to those of the ELISAs. The proposed colloidal gold immunochromatographic method offers advantages such as ease of operability, rapid visualization and high specificity and serves as a practical technical tool for onsite rapid diagnosis and epidemiological surveillance of FAdV-4 infection, thus it has high potential in the prevention and control of HHS.
Keywords: Colloidal gold immunochromatography, Fowl Adenovirus serotype 4, Monoclonal antibody, Penton protein, rapid diagnosis
Received: 01 Dec 2025; Accepted: 06 Feb 2026.
Copyright: © 2026 Luo, Yang, Huang, Wei, Xie, Li, Wu, Ruan, Wang, Zhang, Li, Xie, Yan, Wang and Wei. 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:
Zhixun Xie
Ping Wei
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