Objective To analyze the detections of 15 respiratory pathogens in 4 960 acute respiratory infection (ARI) cases in sentinel hospitals in Guiyang, Guizhou province, from 2024 to 2025, investigate the epidemiological characteristics of each pathogen infection, and provide evidence for the prevention and control of ARI.

Methods Throat swabs were collected from 4 960 ARI patients who visited sentinel surveillance hospitals in Guiyang from 2024 to 2025 for the detections of 15 common respiratory pathogens with multiplex real-time PCR. Software Excel 2021 and SPSS 26.0 were used for the statistical analysis on detection results.

Results In the 4 960 patients, the overall positive rate of pathogen detection was 46.77% (2 320/4 960), and the positive rate of Haemophilus influenzae was highest (11.61%, 576/4 960). In the 2 320 positive samples, the detections of single pathogen accounted for 69.44%（1 611/2 320）, while the detections of multiple pathogens accounted for 30.56%（709/2 320）, the difference was significant (χ²=701.383, P<0.001). The positive detection rate was significantly higher in the outpatient and emergency cases (50.12%, 1 664/3 320) than in the hospitalized cases (40.00%, 656/1 640) (χ²=45.162, P<0.001). The differences in rhinovirus detection rate (χ²=5.361, P=0.021) and group A streptococcus detection rate (χ²=5.395, P=0.020) were significant between men and women. Children and adolescents aged 0 - 18 years were the group at high risk for multi-pathogen infection. There were differences in the detection peak among different pathogens. The total detection rates of pathogens in spring, summer, and winter were significantly higher than those in autumn (χ²=37.959、34.736、37.072, P<0.001). The positive detection rate of total pathogens in Guiyang Municipal Maternal and Child Care Health Center (56.44%, 1 761/3 120) was higher than that in Guizhou People’s Hospital (30.38%, 559/1 840), the difference was significant (χ²=315.772, P<0.001).

Conclusion The detections of ARI pathogens showed significant age and season specific differences, and different pathogens spread alternatively. Haemophilus influenzae, Streptococcus pneumoniae, and influenza virus were predominant respiratory pathogens in Guiyang, and mixed infection was common. The detection rates of the 15 pathogens varied significantly by patient sources. Individuals under 18 years old were identified as a key target group for the prevention and control of ARI.