This paper describes a vacuum-accelerated microfluidic immunoassay (we abbreviate it as VAMI) by sandwiching a filter membrane between a two-layer chip. A direct assay of IgG demonstrated that VAMI could simultaneously achieve higher sensitivity and require less time compared with conventional microfluidic immunoassays. We further applied VAMI to carry out a 3-step competitive assay (including antigen immobilization, competitive reaction and 2(nd) antibody reaction) for detecting the illegal food additive Sudan Red. A total assay time of 15 min with a limit of detection (LOD) of 1 ng ml(-1) is achieved.