BACKGROUND Early diagnosis of osteoarthritis (OA) is difficult. Cartilage oligomeric matrix protein (COMP) and crosslinked C-telopeptides of type II collagen (CTX-II) are two markers which can potentially predict the destruction of articular cartilage of early OA. To comprehensively evaluate the diagnosis value of serum COMP and CTX-II markers in OA, the longitudinal and combined measurement of serum COMP and CTX-II were performed at different stages of pathological process of OA in adult rabbits with OA, compared with the sham-operated rabbits. METHODS Thirty-six adult white rabbits were randomly divided into two groups, the OA group and the control group (n = 18 per group). OA models were established by anterior cruciate ligament transection. Sham operations were performed in the control group. Before the surgery and at weeks 2, 4, 6, 8, 10, and 12 after surgery, serum CTX-II and COMP concentrations were detected using sandwich-ELISA in all rabbits. Three rabbits in each group were killed at weeks 2, 4, 6, 8, 10, and 12 after surgery, and femoral condyle specimens were collected. Histological changes in articular cartilage were evaluated according to the Mankin scoring criteria. RESULTS At each time point, the Mankin scores and serum concentrations of CTX-II and COMP were significantly higher in the OA group than in the control group. In addition, in the OA group, there was a significant relationship between the CTX-II and COMP concentrations and the Mankin score. CONCLUSIONS Early dynamic combined detection of serum CTX-II and COMP concentrations is effective for early OA diagnosis and evaluation of OA severity.