The invention relates to a construction method of highway bridges and particularly relates to an on-beam beam transporting simulation loading test method for a prestressed concrete full-hole prefabricated box beam. The method is applied to the construction of the highway bridges and comprises the following steps: (A) simulating and testing loading conditions of one of two beams by virtue of a single-side static test; (B) simulating the load of a beam transporting vehicle by virtue of a multipoint loading manner, recording parameter differences of the stress state and the actual beam transporting vehicle effect of the tested beam when loads on 5 points/7 points/9 points/11 points are respectively tested, and making comparison to select loading number of points; (C) carrying out finite element simulation on the specific loading working conditions of the tested beam under the loading number of points selected in the step B; and (D) if the stress state in the step C is normal, carrying out an actual load test, so as to detect the stress state, deflection and crack condition of the tested beam on each tested cross section. The on-beam beam transporting simulation loading test method for the prestressed concrete full-hole prefabricated box beam is applicable to the highway bridges.