The potential role of serum RBP4 and THBS2 as biomarker in colorectal cancer (CRC) diagnosis has never been studied. We investigated in large sample using quantitative ELISA method to explore whether serum RBP4 and THBS2 can act as biomarkers for CRC diagnosis. The concentration of RBP4 and THBS2 was measured in 402 CRC patients' serum samples and 218 normal controls' serum samples. The results showed that the average RBP4 and THBS2 concentrations in normal controls were significantly higher than in CRC patients (36.5±11.4μg/mL vs 21.8±8.7μg/mL and 20.5±6.1ng/mL vs 14.5±7.3ng/mL, respectively), both p<0.001. RBP4 distinguished CRC patients from normal individuals with the area under the receiver operating characteristic curve (AUC) performing at 0.852, with sensitivity of 74.9% and specificity of 81.7%. While THBS2 distinguished CRC patients performing AUC at 0.794, with sensitivity of 64.9% and specificity of 87.1%. The ability of RBP4 and THBS2 serum concentration distinguishing CRC from normal controls showed better than that of serum CEA (AUC=0.818) or CA19-9 (AUC=0.650) concentration. This is the first study to report RBP4 and THBS2 as diagnosis serum biomarkers for CRC, which might be a good supplement for CEA or CA19-9 for clinical diagnosis.