ObjectivesThis cross-sectional study aimed to explore job satisfaction among healthcare staff in Guangdong following the health system reforms in 2009, and to investigate the association between job satisfaction and work stress, work–family conflict and doctor–patient relationship.DesignCross-sectional survey.SettingThe Fifth National Health Service Survey was carried out in Guangdong, China.ParticipantsAll participants in this study were healthcare staff including physicians, nurses and public health staff from hospitals, health service centres and health clinics. A total of 6583 questionnaires were distributed and collected. After excluding the incomplete questionnaires, 5845 questionnaires were included for the analysis.Outcome measuresSociodemographic information and scores for evaluating job satisfaction, work stress, work–family conflict and doctor–patient relationship were obtained using the questionnaire developed by the National Health and Family Planning Commission of the People's Republic of China. To assess the significantly associated factors on job satisfaction of the healthcare staff in Guangdong, a binary logistic regression model was used.ResultsBased on the 5845 valid responses of the healthcare staff who worked in Guangdong, the mean score of overall perception of job satisfaction was 3.99 on a scale of 1–6. Among the sociodemographic variables, occupation, educational background, professional status, years of service, annual income and night shift frequency significantly influenced the level of job satisfaction. Work stress, work–family conflict and doctor–patient relationship also had significant effect on job satisfaction.ConclusionsThe overall job satisfaction exceeded slightly dissatisfied (score 3) and approached slightly satisfied (score 4). Measures to enhance job satisfaction include the reduction of workload, increase of welfare, maintaining moderate stress and balancing work–family conflict. Moreover, relevant laws should be issued to protect the healthcare staff from violent acts.