This paper reports an experimental on the flexural performance of prestressed concrete-encased high-strength steel beams (PCEHSSBs). To study the applicability of high-strength steel (HSS) in prestressed concrete-encased steel beams (PCESBs), one simply supported prestressed concrete-encased ordinary-strength steel beam (PCEOSSB) and eight simply supported PCEHSSBs were tested under a four-point bending load. The influence of steel strength grade, I-steel ratio, reinforcement ratio and stirrup ratio on the flexural performance of such members was investigated. The test results show that increasing the I-steel grade and I-steel ratio can significantly improve the bearing capacity of PCESB. Increasing the compressive reinforcement ratio of PCEHSSB can effectively improve its bearing capacity and ductility properties, making full use of the performance of HSS in composite beams. Increasing the hoop ratio has a small improvement on the load capacity of the test beams; setting up shear connectors can improve the ductile properties of the specimens although it does not lead to a significant increase in the load capacity of the combined beams. Then, combined with the test data, the comprehensive reinforcement index considering the location of reinforcement was proposed to evaluate the crack resistance of specimens. The relationship between the comprehensive reinforcement index and the crack resistance of specimens was given.