Development and diagnostic validation of a recombinant Hexon L1-based indirect enzyme-linked immunosorbent assay for detecting antibodies to fowl adenovirus serotype 8b in chickens.
5 affiliations
- 1Department of Farm Resources and Animal Production, Faculty of Veterinary Medicine, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom, 73140, Thailand.
- 2Animal Health and Biomedical Sciences Study Program, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, 10900, Thailand.
- 3Department of Pathology, Faculty of Veterinary Medicine, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom, 73140, Thailand.
- 4Kamphaeng Saen Veterinary Diagnostic Center, Faculty of Veterinary Medicine, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom, 73140, Thailand.
- 5Department of Pathology, Faculty of Veterinary Medicine, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom, 73140, Thailand; Center for Advanced Studies for Agriculture and Food, Kasetsart University Institute for Advanced Studies (KUIAS), Kasetsart University, Bangkok, 10900, Thailand.
Abstract
Inclusion body hepatitis (IBH), caused by fowl adenoviruses (FAdVs), is a significant disease impacting poultry productivity. Recent data indicates that FAdV serotype 8b (FAdV-8b) is the predominant serotype associated with IBH cases in Thailand. Although the serum neutralization (SN) test is a commonly used method for detecting serotype-specific neutralizing antibodies, it has limitations in its routine application. This study developed and diagnostically validated an FAdV-8b-targeted indirect ELISA (iELISA) utilizing a recombinant protein of the FAdV-8b Hexon loop-1 region (rHexon8b), which was generated in E. coli and purified via Ni-NTA affinity chromatography. A total of 467 chicken sera with validated SN results were utilized for test optimization and diagnostic assessment. The optical density measured at 450 nm (OD₄₅₀) cut-off value was set at 0.112 and was supported by receiver operating characteristic analysis, which demonstrated excellent discriminatory performance with an area under the curve of 0.96 (95% confidence interval, 0.9407-0.9748; p < 0.0001). Utilizing the SN test as the reference standard, the rHexon8b iELISA demonstrated a diagnostic sensitivity of 92.73% and a diagnostic specificity of 89.33%, exhibiting substantial concordance between the two assays (Cohen's κ = 0.818). The assay showed no detectable cross-reactivity with the tested sera positive for major chicken pathogens or heterologous FAdV serotypes, including FAdV-2 and -11. The intra- and inter-assay coefficients of variation ranged from 0.92% to 6.80% and from 3.43% to 9.46%, respectively, indicating excellent repeatability and precision. These findings suggest that the rHexon8b iELISA is a rapid, reproducible, and practical serological screening tool for flock-level surveillance, vaccination monitoring, and epidemiological investigation of FAdV-8b-associated IBH in chickens.
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BibTeX
@article{kulprasertsri2026development,
title = {{Development and diagnostic validation of a recombinant Hexon L1-based indirect enzyme-linked immunosorbent assay for detecting antibodies to fowl adenovirus serotype 8b in chickens.}},
author = {Sittinee Kulprasertsri and Malee Thongnid and Ploypassorn Homklinkaew and Siriluk Jala and Sakuna Phatthanakunanan and Preeda Lertwatcharasarakul and Thaweesak Songserm},
journal = {Poultry science},
year = {2026},
month = sep,
doi = {10.1016/j.psj.2026.107864},
url = {https://europepmc.org/article/MED/42833010},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Development and diagnostic validation of a recombinant Hexon L1-based indirect enzyme-linked immunosorbent assay for detecting antibodies to fowl adenovirus serotype 8b in chickens.
AU - Sittinee Kulprasertsri
AU - Malee Thongnid
AU - Ploypassorn Homklinkaew
AU - Siriluk Jala
AU - Sakuna Phatthanakunanan
AU - Preeda Lertwatcharasarakul
AU - Thaweesak Songserm
PY - 2026
DA - 2026/09/30
JO - Poultry science
DO - 10.1016/j.psj.2026.107864
UR - https://europepmc.org/article/MED/42833010
AB - Inclusion body hepatitis (IBH), caused by fowl adenoviruses (FAdVs), is a significant disease impacting poultry productivity. Recent data indicates that FAdV serotype 8b (FAdV-8b) is the predominant serotype associated with IBH cases in Thailand. Although the serum neutralization (SN) test is a commonly used method for detecting serotype-specific neutralizing antibodies, it has limitations in its routine application. This study developed and diagnostically validated an FAdV-8b-targeted indirect ELISA (iELISA) utilizing a recombinant protein of the FAdV-8b Hexon loop-1 region (rHexon8b), which was generated in E. coli and purified via Ni-NTA affinity chromatography. A total of 467 chicken sera with validated SN results were utilized for test optimization and diagnostic assessment. The optical density measured at 450 nm (OD₄₅₀) cut-off value was set at 0.112 and was supported by receiver operating characteristic analysis, which demonstrated excellent discriminatory performance with an area under the curve of 0.96 (95% confidence interval, 0.9407-0.9748; p < 0.0001). Utilizing the SN test as the reference standard, the rHexon8b iELISA demonstrated a diagnostic sensitivity of 92.73% and a diagnostic specificity of 89.33%, exhibiting substantial concordance between the two assays (Cohen's κ = 0.818). The assay showed no detectable cross-reactivity with the tested sera positive for major chicken pathogens or heterologous FAdV serotypes, including FAdV-2 and -11. The intra- and inter-assay coefficients of variation ranged from 0.92% to 6.80% and from 3.43% to 9.46%, respectively, indicating excellent repeatability and precision. These findings suggest that the rHexon8b iELISA is a rapid, reproducible, and practical serological screening tool for flock-level surveillance, vaccination monitoring, and epidemiological investigation of FAdV-8b-associated IBH in chickens.
SN - 0032-5791
SN - 1525-3171
ER -