ArticlePoultry science2026
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.
Article in Poultry science, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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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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