Objective · To design an immuno-affinity chromatography device for the separation and detection of syphilis specific antibody, then verify its performance of detection and clinical application. Methods · Affinity filler packed by Treponema pallidum (TP) antigen in affinity chromatography can specifically adsorb TP specific antibody (including IgG and IgM) in samples. After balance, elution and desalination, IgG or IgM gold labeled chromatography strip detects the possibly present syphilis specific IgG or IgM antibody. Twenty cases of syphilis antibodynegative samples and 230 cases of syphilis antibody positive clinical specimens were detected by this chromatography device, and 40 cases were also detected by Western blotting.Results ·The standard operation procedure of the affinity chromatography device was optimized, which could effectively detect the specific IgG and IgM antibody of syphilis. The results of 20 syphilis antibody negative samples were all negative. In 230 syphilis antibody positive cases, the detection results were 2 cases with TP-IgG(-) and TP-IgM(-), 210 cases with TP-IgG(+) and TP-IgM(-),10 cases with TP-IgG(-) and TP-IgM(+), and 2 cases with TPIgG(+)and TP-IgM(+). The detection results of 40 cases were compared with the results detected by Western blotting, among which 2 cases detected by affinity chromatography device were TP-IgG(-) and TP-IgM(-), while the results detected by Western blotting were TP-IgG(+) and TP-IgM(-). But the results of the two methods showed no statistically significant difference (P>0.05). Conclusion · The application of the device in separation and detection of IgG and IgM antibodies against TP pathogens is feasible, and it has important value for further application in clinical diagnosis.