Abstract： Objective To investigate the role of and relationship between reactivity to autologous serum and dermatophagoides farinae (Df) in the pathogenesis of chronic urticaria (CU). Methods Autologous serum skin test (ASST) and skin prick test (SPT) to Df were carried out in 831 patients with CU. The correlation between reactivity to autologous serum and Df was statistically analyzed. Results The positivity rate of ASST and SPT to Df was 51.74% and 64.62%, respectively in the 831 patients. SPT was positive in 56.52% of patients with positive ASST and in 73.86% of those with negative ASST (P ＜ 0.05). In patients with positive ASST, the degree of autologous serum reactivity was negatively correlated with that of reactivity to Df (P ＜ 0.05). Conclusions The skin reactivity to Df and autologous serum plays an important role in the pathogenesis of CU, and the degree of the reactivity to Df and autologous serum is negatively correlated. To conduct ASST and SPT simultaneously in patients with CU may favor the clinical classification and therapy of CU.