Interferons (IFNs) play a role in inhibition of tumor growth and participate in immunoreactions. Among IFNs, interferon-gamma (IFN gamma) is one of the most important therapeutic proteins and its immunodulation ability is better than that of other types. The objective of this study is to develop a manual self-assembled colloidal gold nanoparticle-immunochromatographic strip for human IFNg using anti-human IFNg polyclonal and monoclonal antibodies. Colloidal gold with a 25 nm diameter was made from chloroauric acid (HAuCl4), and labeled on anti-IFN gamma mAbs as a chrominance reagent. A good linear relationship existed between the pixel intensity and the human IFNg concentrations from 10-1000 ng mL(-1) in mouse serum and buffer, respectively, the regression equation was Y = 0.159logX + 0.0648, R-2 = 0.992 in mouse serum; Y = 0.294logX + 0.091, R-2 = 0.9969 in phosphate buffer by this proposed strip. Moreover, in the determination for mouse serum samples no cross-reaction occurred and the detection time was approximately 10 minutes. The shelf life of the strip was above 28 days at room temperature. The major advantages of the manual operation model were no expensive instruments and less reagents required. This proposed strip was highly specific, economic, convenient, and no machine was needed in clinical diagnosis.