The invention discloses an MICA extracellular region mutant, a screening method thereof, an scFv-MICA fusion antibody, and a preparation method and application of the scFv-MICA fusion antibody. The MICA extracellular region mutant is mutated at the 24th site, the 33th site, the 69th site, the 112th site and the 126th site. The screening method comprises the following steps: constructing a mutant MICA bacteriophage library, expressing recombinant protein NKG2D-Fc, screening bacteriophage and the like. The scFv-MICA fusion antibody is formed by linking scFv and an MICA mutant through a flexiblepeptide. The preparation method comprises the following steps: constructing a recombinant gene of a fusion antibody, constructing a recombinant vector, transfecting competent cells, performing expressing and purifying, and the like. The MICA extracellular region mutant disclosed by the invention can be used for efficiently inducing the cytotoxic effect of NKG2D mediated NK cells; the fusion antibody constructed by the MICA extracellular region mutant and scFv can utilize a tumor specific antibody part to efficiently target tumor cells, and meanwhile, the high-affinity mutation MICA part can mediate NK cells and the like to effectively kill target cells.