The present invention, specifically, the sample pad than to a kit for the simultaneous detection of multiple disease using a highly sensitive detection method and simultaneous three-dimensional protein nanoparticle probe of high sensitivity 3D protein nanoparticle probes based on multiple disease; A signal molecule ingredients conjugated pad; Self-assembly (self-assembly) the membrane comprises a protein nanoparticle probe, which is a disease marker specific epitope fusion protein expressed on the surface; And as for the simultaneous detection of multiple disease method using a lateral flow assay kit, and the kit comprising an absorbent pad, a protein nanoparticle probes according to the present invention are specific epitopes on the surface of disease markers have the correct orientation, the high-density expression is possible, by adjusting their mixing ratio of which can detect a specific epitope several types of disease markers in various amounts, this detecting method of using the disease marker on the surface of a specific epitope various disease marker protein nanoparticles it is expressed, the sensitivity and accuracy is very excellent.