The invention discloses a TMV-CMV-PVY three-virus (tobacco mosaic virus, cucumber mosaic virus and potato virus) colloidal gold rapid test strip, which is composed of a sample pad, a colloidal gold pad, an NC membrane, a piece of water-absorbent filter paper and a backing, wherein a colloidal gold labeled TMV specific antibody, a CMV specific antibody and a PVY specific antibody are enveloped on the colloidal gold pad, and the three antibodies are secreted from hybridoma cell strains BALB/c 15 50, BALBc 15 27 and BALBc 15 8; a detection line and a control line are arranged on the NC membrane; specific antigens of three viruses are enveloped on the detection line; and a colloidal gold labeled second antibody is enveloped on the control line. The test strip is especially suitable for detecting the TMV, CMV and PVY in Guangdong tobacco-growing areas, and the test strip is rapid, sensitive, accurate, low in cost and convenient to operate; the test strip is capable of simultaneously diagnosing various viruses by conducting sampling once; the test strip is quite suitable for in-situ preliminary screening of a great batch of samples; and the test strip, in actual production, has a high application value, and the test strip has a good popularization and application prospect.