Lateral flow immunochromatographic strip tests (IST) are very promising for use as on-site test diagnostic tools in developing countries.In this work, a fabrication process simplified, lateral flow immunochromatographic test strip for aflatoxin M1 (AFM1) detection in milk samples has been developed.The immunostrip was specific to AFM1 showing no cross-reactivity with other major aflatoxins.The visual detection limit (VDL) and threshold level for AFM1 was 0.3 and 4.0 ng/mL, respectively.Evaluation results of the IST revealed good accuracy and repeatability.In the detection of AFM1 contamination level in milk samples, visual results of the IST are in good correlation with those of a quantitative enzyme-linked immunosorbent assay (ELISA).This AFM1-IST, employing a onestep method, was environmentally friendly without the use of any organic reagents or buffers and additional equipment, and could serve as an on-site and home-use device for AFM1 tests in milk, and be helpful to peoples health care.