Abstract: Objective To analyze the epidemiological characteristics of 21 respiratory pathogens in influenza-like illness (ILI) populations in Jing'an District, Shanghai in 2024, and to provide a scientific basis for the prevention and control of respiratory infectious diseases. Methods Data of 1 907 ILI cases at four sentinel hospitals in Jing'an District were collected from January to December 2024. Nasopharyngeal swab samples were collected and tested for 21 respiratory pathogens using polymerase chain reaction (PCR) methods. Chi-square test and Cochran-Armitage trend test were used for data analyses. Results Among the 1 907 ILI cases, 1340 were tested positive (70.27%), including 1 160 (60.83%) virus-positive cases, 424 (22.23%) bacteria-positive cases, and 86 (4.51%) positive cases of other pathogens (fungi, mycoplasma, and chlamydia). The top five viruses by detection rate were: influenza virus (14.84%), SARS-CoV-2 (14.47%), rhinovirus (12.69%), adenovirus (7.08%), and parainfluenza virus (6.71%). The top two bacteria by detection rate were Streptococcus pneumoniae (14.47%) and Haemophilus influenzae (10.33%). Among other pathogens (fungi, mycoplasma, and chlamydia) Mycoplasma pneumoniae showed the highest detection rate (4.30%). In terms of age distribution, statistically significant differences were observed in the detection rates of SARS-CoV-2 and Klebsiella pneumoniae (P<0.05), with the highest rates found in individuals aged 65 years and above. Statistically significant differences were also found in the detection rates of rhinovirus, adenovirus, enterovirus, common coronavirus, respiratory syncytial virus, bocavirus, parainfluenza virus, human metapneu-movirus, Streptococcus pneumoniae, Haemophilus influenzae, and Mycoplasma pneumoniae among different age groups (P<0.05), all showing the highest detection rates in the 0-＜15 years age group. In terms of seasonal distribution, novel coronavirus, adenovirus, parainfluenza virus, enterovirus, Streptococcus pneumoniae, Haemophilus influenzae, and Mycoplasma pneumoniae showed epidemic peaks in summer; rhinovirus, common coronavirus, and Klebsiella pneumoniae had higher detection rates in autumn;Influenza virus exhibits a peak incidence during winter, while human metapenu-movirus peaks in late winter and spring. Co-infection with two or more pathogens were detected in 409 cases, with a co-infection rate of 30.52%. The 0-＜15 years age group accounted for the highest proportion (88.26%). Conclusion In 2024, the main pathogens of ILI cases in Jing'an District, Shanghai were influenza virus, SARS-CoV-2, rhinovirus, Streptococcus pneumoniae and Haemophilus influenzae. Notably, pathogen detection rates varied by age and season, and co-infection was relatively common.