Objective:To establish a common method for detecting serotypes of respiratory adenovirus, and to detect the main serotypes of respiratory human adenovirus (HAdV) infection in children in Wenzhou area.Methods:A multiplex PCR method based on capillary electrophoresis was developed to detect 12 common serotypes of respiratory adenovirus.A total of 1 059 children with acute respiratory infection who were admitted to Yuying Children′s Hospital Affiliated to Wenzhou Medical University from January 2018 to December 2019 with positive infection of HAdV detected by the direct immunofluorescence method were recruited and retrospectively analyzed.Multiplex PCR was performed to determine 12 serotypes of respiratory adenovirus, including HAdV-1, 2, 3, 4, 5, 7, 14, 21, 37, 40, 41 and 55.Meanwhile, some samples were randomly selected to examine the consistency in the detection result by the first-generation sequencing method.Results:A total of 1 059 specimens of respiratory secretions with positive HAdV antigen were collected.Detected by multiplex PCR method, 947 cases (89.4%) were positive for 1 serotype, 13 cases (1.2%) were mixed infection with 2 serotypes, and 24 cases (2.3%) were negative.In addition, 75 cases(7.1%) were positive but could not be serotyped.Among the 947 children with the positive infection of a single serotype, 415 cases (43.8%) were HAdV-3 in subgroup B, 318 cases(33.6%) were HAdV-7, 12 cases (1.2%) were HAdV-55, 2 cases (0.2%) were HAdV-21, 108 cases (11.4%) were HAdV-2 in subgroup C, 70 cases (7.4%) were HAdV-1, 16 cases(1.7%) were HAdV-5, and 6 cases(0.6%) were HAdV-4 in subgroup E. HAdV-14, HAdV-37, HAdV-40 and HAdV-41 were not detected.A total of 51 positive samples of HAdV infection detected by multiplex PCR were randomly selected to compare with the detection result by the first-generation sequencing, which were all consistent.Conclusions:This study successfully established a multiplex PCR based on capillary electrophoresis in diagnosing common serotypes of respiratory adenovirus infection in children.HAdV-3, HAdV-7 of subgroup B and HAdV-2 and HAdV-1 of subgroup C were the main serotypes of respiratory adenovirus infection in children of Wenzhou area.HAdV-14, HAdV-37, HAdV-40 and HAdV- 41 were not detected.