Purpose: This study was performed to develop a lateral flow strip sensor for the detection of pathogenic Escherichia coli O157:H7 in various samples. Also, feasibility of using an image analysis method to improve the interpretation of the strip sensor was evaluated. Methods: The lateral flow strip sensor has been fabricated based on nitrocellulose lateral-flow membrane. Colloidal gold and E. coli O157:H7 antibodies were used as a tag and a receptor, respectively. Manually spotted E. coli O157:H7 antibody and anti-mouse antibody on nitrocellulose membrane were used as test and control dots,respectively. Feasibility of the lateral flow strip sensor to detect E. coli O157:H7 were evaluated with serially diluted E. coli O157:H7 cells in PBS or food samples. Test results of the lateral flow strip sensor were measured with an image analysis method. Results: The intensity of the test dot started to increase with higher concentration of the cells were introduced. The sensitivities of the sensor were both 104 CFU/mL Escherichia coli O157:H7 spiked in PBS and in chicken meat extract,respectively. Conclusions: The lateral flow strip sensor and image analysis method could detect E. coli O157:H7 in 20 min,which is significantly quicker than conventional plate counting method.