Objective: The objective of the present study was to develop a method for sampling and detecting an adenovirus-derived gene therapy (GT) vector on isolator worksurfaces. Methods: We used a quantitative PCR (q-PCR) to detect the viral genome in standard dilutions of pure GT product and extracts of sampled surfaces. We compared three devices for surface sampling (a cotton compress, a cotton swab and a polyester flocked swab) and performed positive control, negative control and induced contamination tests for each. Results: Our results showed that the GT pure product is detected by the q-PCR assay and is amplified throughout the range of dilutions. The mean difference between the expected and measured number of vector particles in the q-PCR assay was 1.27 log. The numbers of particles in the total extracted volume were 4.66×108 for the polyester swab (7.8% of the initial quantity), 3.82×108 for a cotton compress (6.4%) and 2.88×107 for a cotton swab (4.8%). Conclusion: These initial results suggest that viral monitoring of worksurfaces is feasible and will help us to validate the GT product supply chain.