In order to accurately calculate the long-term prestress losses of prestressed tendons, a time-varying model of long-term prestress loss considering the interaction between concrete shrinkage, creep, and the stress relaxation of prestressed tendons was constructed. Then, a method for calculating the long-term prestress losses of concrete structures was developed. A long-term prestress loss test of a prestressed concrete T-beam in a long-term field test environment was carried out. The measured values of long-term prestress losses are compared with the calculated results of JTG 3362-2018, AASHTO LRFD-2007, and the time-varying law model. The results show that the long-term effective tension of the T-beam decreases gradually with the increase in the load holding time. At the beginning of loading, the tensile force changes rapidly and then gradually slows down. The later the tensile age or the higher the initial loading stress level, the smaller the long-term prestress losses of the prestressed tendons. The long-term prestress loss values calculated by JTG 3362-2018, AASHTO LRFD-2007, and the time-varying law model increase with the increase in the load holding time. In the early stage of loading, the rate of change slows down and tends to be stable. The calculated results of JTG 3362-2018 and AASHTO LRFD-2007 are significantly different from the measured values. However, the calculated results of the time-varying law model are in good agreement with the measured values. The average coefficients of variation of the long-term prestress loss calculated by JTG 3362-2018, AASHTO LRFD-2007, and the time-varying law model are 17%, 10%, and 5%, respectively. The time-varying law model of the long-term prestress losses of prestressed tendons is accurate, and the long-term prestress loss of prestressed reinforcement can be predicted effectively.