Objective To understand the pathogen detection of inpatients in a hospital before antimicrobial treatment, take comprehensive intervention measures, and improve pathogen detection rate before antimicrobial treatment in this hospital. Methods Patients who received antimicrobial treatment from October to December of 2021 in a tertiary first-class hospital in Changzhi City of Shanxi Province were selected as the control group, comprehensive intervention measures were taken from January 1, 2022, patients who received antimicrobial treatment from April to June of 2022 were selected as the intervention group. Pathogen detection rate before antimicrobial treatment, pathogen detection rate before antimicrobial treatment in key departments, detection rate of sterile specimens and detection rate of important multidrug-resistant organisms between two groups were compared. Results Pathogen detection rate before treatment in the intervention group was higher than that in the control group (53.06% vs 30.61%, P < 0.01). Detection rate in 5 key departments after intervention was improved, difference was statistically different (all P < 0.05). Detection rate of sterile specimens in the intervention group was higher than that in the control group (15.58% vs 7.52%, P < 0.01). Detection rate of carbapenem-resistant Acinetobacter baumannii in the intervention group was significantly lower than that in the control group (64.94% vs 84.00%, P < 0.01). Detection rates of methicillin-resistant Staphylococcus aureus and carbapenem-resistant Pseudomonas aeruginosa in the intervention group were not significantly different from those in the control group (both P>0.05). Conclusion The comprehensive intervention measures can improve the pathogen detection rate and the sterile specimen detection rate of inpatients in this hospital before antimicrobial treatment, but fail to effectively reduce the detection rate of important multidrug-resistant organism, thus need to be improved.