Disclosed are a battery isolating strengthening membrane and a preparation method thereof. The battery isolating strengthening membrane is characterized by being prepared from a biaxially oriented expanded polytetrafluoroethylene microfiltration membrane, which achieves a longitudinal tensile strength of 25-30 Mpa, a transverse tensile strength of 20-25 Mpa, a longitudinal elasticity modulus of 70-80 Mpa, a transverse elasticity modulus of 50-60 Mpa, and longitudinal and transverse maximum force of 3-4 N. The preparation method of the battery isolating strengthening membrane comprises the steps of material blending, pushing, extrusion, rolling, lubricant removal, longitudinal orientation, thermal treatment, cooling and shaping, transverse orientation, thermal formation and cooling formation. The battery isolating strengthening membrane is prepared through reasonable processes of multiple orientation, thermal treatment at temperature higher than normal melting point, and cooling; the prepared battery isolating strengthening membrane is an orderly arranged crystallized structural membrane, which is low in knotting, high in tensile strength and elasticity modulus, and good in creep resistance, hardness and dimensional stability and the like. With longitudinal and transverse strengths closely similar, the prepared battery isolating strengthening membrane can not only be better in dimensional stability and higher in mechanical performance, but also significantly reduce battery internal resistance to improve the battery performance and reduce membrane material consumption and thecost.