The invention provides a method for comprehensively evaluating the titer of adenovirus pAd-M3C by combining TCID50, MOI, GTU and PFU. Three detection methods of TCID50, MOI and GTU are integrated, after experimental operation steps are improved and refined, the adenovirus pAd-M3C titer is detected at the same time, conversion and comparison are carried out through PFU, and the TMGP method is called for short, so that an original single method can be optimized and improved in repeatability, stability, sensitivity, usability and time consumption, the effectiveness of adenovirus titer detection is improved, and the method is suitable for popularization and application. And a foundation is laid for subsequent experimental standard operation. The TMGP method can be used for estimating the MOI value of the virus titer within 24-48 hours, verifying GTU data within 36-48 hours and comparing the accurate titer of TCID50 on the most critical tenth day, so that repeated experiments are reduced, the experiment cost is reduced, and the effectiveness is improved.