OBJECTIVE: To identify factors associated with acceptance of the pandemic Influenza A H1N1 California 7/09 vaccine (pH1N1 vaccine) in rural hospital staff. DESIGN: Cross-sectional group administered survey. SETTING: A 58-bed rural Queensland hospital, over a 10-day period starting five days after commencement of the pH1N1 vaccination program. PARTICIPANTS: Hospital staff and students. MAIN OUTCOME MEASURES: Intention to receive pandemic influenza vaccination, seasonal influenza vaccination. RESULTS: The survey was completed by 252 staff and students (full response rate 76.1%, cooperation rate 98.4%). The majority of respondents had already decided whether or not to receive the pH1N1 vaccine: 111 (44.0%) intended to be vaccinated, 63 (25.0%) intended not to be vaccinated and 78 (31.0%) were undecided. Of 125 staff who had received the seasonal influenza vaccine, 86 (68.8%) intended to receive the pandemic vaccine. Respondents were less likely to state acceptance of the pH1N1 vaccine if they identified concern about vaccine adverse reactions (odds ratio (OR) 0.29; 95% confidence interval (CI), 0.11-0.78), vaccine safety (OR 0.20; 95% CI, 0.09-0.46), vaccine efficacy (OR 0.20; 95% CI, 0.04-0.87) or perceived limitations of vaccine trials (OR 0.08; 95% CI, 0.01-0.99) as primary reasons discouraging pandemic influenza vaccination. CONCLUSIONS: There is considerable concern among rural hospital staff regarding pH1N1 vaccine adverse reactions, safety, efficacy and the adequacy of clinical trials on its use. Education specifically addressing these concerns might increase pandemic vaccine acceptability.