The invention provides a preparation method of a graphene conductive structure body. By employing methods of repeated coating and drying of a graphene solution and an isolation solution in order, multiple graphene layers and multiple isolation layers which are arranged at intervals are formed at the outer layer/the upper layer of a base material inner layer so as to rapidly and simply prepare a graphene conductive structure body, and the prepared graphene conductive structure body is high in conductivity and high in conduction efficiency, and meanwhile, in order to prevent a certain layer or multiple layers of single graphene films from being broken to cause non-conductivity, a conductive pile position structure is creatively designed on a single isolation insulating film, and the two isolated layers of single graphene films are connected in a short circuit manner to ensure the conductivity.