Gene Human leukocyte antigen (HLA)-A3101 is known responsible of a medicine allergy which may cause a fatal outcome. General clinics need a quick test to determine if a patient is engaged with the related gene defect and avoid the medical prescription from suspicious drugs. In this study, Gene HLA-A3101 is detected on membrane-based lateral-flow (MBFL) strips, incorporated with an immune reaction and color readouts developed by gold nanoparticles. The HLA-A-3101 DNA modified with biotin and digoxigenin labels was amplified by the Polymerase Chain Reaction (PCR) and then detected on the MBFL strips. The optimal condition of antibody concentration was at least 10µg/ml, to avoid the antibody-gold complex from aggregation. The signal can be observed as low as 0.49 ng of the PCR product by naked eyes. The specificity of PCR primers was also investigated their annealing validity. The detection specificity from other species genes, including a plant, a virus, and a human gene, were also verified. The coefficient of variation values of the intra assay, which tests the same sample in the same day, and the inter assay, which tests three samples in three consecutive days, are in satisfactory ranges of 4.6% and 6.5%, respectively.