The invention discloses a conductive particle and a preparation method thereof, and an anisotropic conductive film. The preparation method of the conductive particles comprises the following steps: bytaking a silicon-containing precursor as a silicon source and water and alcohol as a mixed solvent, performng stirring, centrifuging and drying to obtain mesoporous silica spheres; performing stirring, curing and calcining by taking the mesoporous silica spheres as a template, a resin solution as a carbon source and water and alcohol as a mixed solvent to obtain porous carbon spheres; dissolvinganiline and the porous carbon spheres in an acidic solution, adding an initiator, performing washing and drying to obtain a porous carbon sphere polyaniline nanowire compound which is conductive particles. According to the technical scheme, the problems that existing conductive particles are higher in cost, poorer in compatibility with organic matter, larger in particle size and higher in materialcost due to the fact that the needed filling amount is large in the actual application process can be solved.