Objective: To establish a high-throughput chemiluminescence assay of serotype 5 specific neutralizing antibody and understand the epidemiology of this antibody in the healthy adults and children in Guangzhou.

Methods: Using rAd5 carrying the reporter gene of secreted alkaline phosphatases (SEAP), serum samples from 116 healthy adults and 94 healthy children were examined with chemiluminescence assay to detect the presence of Ad5 neutralizing antibody. The reliability of this assay was tested against conventional cytopathic effect observation.

Results: The chemiluminescence assay using secreted alkaline phosphatases (SEAP) as the reporter allowed rapid, sensitive, specific and reproducible detection of serotype 5 specific neutralizing antibody for epidemiological study of Ad5, which was positive in 26.72% of the adults and 17.02% of the children in this study.

Conclusion: Ad5 neutralizing antibody is prevalent in the population of Guangzhou, suggesting the necessity of developing other serotype adenovectors for better vaccination and therapeutic effects.

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