This study identified the prevalence of and risk factors for non-compliance with glove utilization and hand hygiene among rural obstetrics and gynaecology workers in two poor counties in Anhui, China. A cross-sectional survey was conducted using a self-administered anonymous semi-structured questionnaire. The study population included all health workers in the departments of obstetrics and gynaecology in township hospitals and county health facilities. Among the 137 participants, the rate of non-compliance with glove utilization was 61%, the rate of non-compliance with hand hygiene was 40%, and the rate of non-compliance with both was 67%. 'Unnecessary and inconvenient' accounted for 66% of the reasons for not always using gloves and 71% for not always washing hands. 'Short of gloves or water' accounted for 15% and 22% of reasons, respectively. Two variables were associated with non-compliance: county [odds ratio (OR)=11.56, P<0.001] and knowledge (OR=0.10, P=0.033). After removing county from the regression model, institutional level (OR=2.34, P=0.059), institutional support (OR=3.85, P=0.009) and knowledge (OR=0.05, P=0.007) were associated with non-compliance. The prevalence of non-compliance with glove utilization and hand hygiene was high. The risk factors for non-compliance were institutional level, institutional support and knowledge. These variables incorporated both subjective and objective reasons for non-compliance.