Objective: To analyze the detection spectrum of specific multidrug-resistant bacteria, the incidence of hospital infections, and the characteristics of institutional stratification in monitored medical institutions in a certain city, so as to provide a basis for regional differentiated prevention and control. Methods: The regional platform data of 13 surveillance medical institutions in a certain city in 2025 were retrospectively collected, and indicators such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), carbapenem-resistant Enterobacteriaceae (CRE), carbapenem-resistant Acinetobacter baumannii (CRAB), and carbapenem-resistant Pseudomonas aeruginosa (CRPA) were included. Stratified according to hospital grade and bed size, detection rate, absolute difference, range, Z-score standardization, and Ward hierarchical clustering analysis were used. Results: The overall detection rate of multidrug-resistant bacteria in the city was 6.25%; CRAB, MRSA, and CRPA were 29.87%, 18.91%, and 14.69%, respectively, and the remaining bacterial species were all less than 5%. The overall detection rates of tertiary and secondary hospitals are similar, but the CRAB of secondary hospitals is higher, and the CRPA of tertiary hospitals is higher. Institutions with 501 to 1,000 beds have the highest overall detection rate, while institutions with ≤ 300 beds have the lowest rates, forming a relatively independent low detection spectrum. The city's hospital infection incidence rate and case incidence rate of multidrug-resistant bacteria are both 0.02%. Conclusion: Regionally specific multidrug-resistant bacteria are heterogeneous in species and institutional levels. CRAB, MRSA, and CRPA are key risk bacteria. Stratified early warning and precise prevention and control should be implemented based on bacterial species spectrum, institutional level, and change amplitude.