The aim of this cross-sectional study was to explore the association of worker characteristics and work organization factors with prevalence of patient-handling injury among nursing personnel in an acute-care inpatient setting. Self-administered questionnaires (n = 585) captured worker characteristics and patient-handling injuries within the previous 6 months. Karasek's Job Content Questionnaire measured work organization factors, including job strain (high psychological demand, low decision latitude). We created a novel measure (job strain(PHYSICAL)) reflecting high physical demand and low decision latitude, providing a more direct physiologic link to our outcome. Log-binomial regression was used to calculate prevalence ratios (PR) and 95% confidence intervals (CI). Patient-handling injuries were prevalent (35%), and incident reports were filed infrequently for injuries receiving medical care. Prevalence of injury was higher among aides compared to nurses [adjusted PR 1.4, 95%CI (1.1-1.8)] as well as among those with high [adjusted PR 1.6, 95%CI (1.2-2.4)] or mid [adjusted PR 1.9, 95%CI (1.4-2.7)] levels of job dissatisfaction. The novel definition of job strain (job strain(PHYSICAL): high physical demand, low decision latitude) was more strongly associated with patient-handling injury than the traditional definition of job strain (high psychological demand, low decision latitude). These findings add to a growing body of literature on the highly contextual nature of work organization factors.