Prestressed concrete Box girder bridge has been widely used in modern long-span bridge structures because of its good overall structural mechanical performance. However, in the concrete Box girder bridge built so far, the phenomenon of box girder cracking and excessive downward deflection during the operation stage is common. The lack of understanding of concrete shrinkage and creep effect and its effect in actual concrete Box girder bridge is one of the possible reasons. The current calculation formulas for concrete shrinkage and creep are mostly based on the results of laboratory model tests. Due to the large size of actual concrete box girder structures and their complex natural environment, it is particularly important to conduct long-term testing on the actual structure to obtain measured data that can verify the current specifications for concrete shrinkage and creep calculation formulas. Combined with the construction and operation of two large-span prestressed concrete Box girder bridge on an expressway, the real response of Box girder bridge in natural environment under the action of concrete shrinkage and creep is tested, and the test data are analyzed in detail. On this basis, a calculation method of concrete shrinkage strain and creep coefficient is proposed, taking into account the factors such as concrete temperature, environmental relative humidity, local theoretical thickness of box girder and their changes, And it is applied to the analysis of shrinkage and creep effect of actual bridges, and some conclusions with practical value are obtained, which provide reference for the calculation of shrinkage and creep of actual Box girder bridge