The invention relates to a targeted superparamagnetic monodisperse nano-flower probe, the preparation and the application thereof. A nano-flower probe is composed of three parts, namely a magnetic gold-bearing particle nano-flower, a hyperbranched macromolecular HPG and a target molecule, wherein one end of the hyperbranched macromolecular HPG is provided with a mercapto group and a branched terminal. The nano-flower probe is prepared through the hot solvent method. That is, firstly, a Fe3O4 carrier is prepared. Secondly, a golden seed is loaded on the carrier, and then the magnetic gold-bearing particle nano-flower is obtained. Finally, the magnetic gold-bearing particle nano-flower is connected sequentially with the HPG and the target molecule. The above nano-probe is applied to the oncotherapy for animal in-vivo double-targeted imaging under the alternating magnetic field. Compared with the prior art, the targeted superparamagnetic monodisperse nano-flower probe is simple to prepare, low in cost, high in targeting efficiency, and small in oncotherapy side effect. Compared with the prior art, the targeted superparamagnetic monodisperse nano-flower probe has the advantages of simple preparation, low cost, high targeting efficiency, small oncotherapy side effect and the like.