The organic Rankine cycle (ORC) is a popular technology used in waste heat recovery and medium-low-temperature heat utilization. Working fluid plays a very important role in ORC. The selection of working fluid can greatly affect the efficiency, the operation condition, the impact on the environment, and the economic feasibility of ORC. The expander is a key device in ORC. As a novel expander, single-screw expanders have been becoming a research focus in the above two areas because of their many good characteristics. One of the advantages of single-screw configurations is that they can conduct a vapor–liquid two-phase expansion. Therefore, in order to give full play to this advantage, a working fluid selection for ORC using a single-screw expander was conducted in this paper. Three indicators, namely, net work output, thermal efficiency, and heat exchange load of condenser, were used to analyze the performance of an ORC system. Through calculation and analysis, it can be seen that an ORC system that uses a single-screw expander and undergoes a vapor–liquid two-phase expansion is able to obtain a higher thermal efficiency, higher net work output, and a smaller heat exchange load of the condenser. Regardless of whether isentropic efficiency of the expander is considered or not, cis-butene may be the best candidate for working in subcritical cycles. HFO working fluids are more suitable for working in transcritical cycles, and HFO-1234ze(E) may be the best.