ObjectiveTo understand microbial contamination and existing problems of B ultrasonic probe with disinfectant coupling agent, and prevent healthcare associated infection caused by B ultrasonic probe.MethodsB ultrasonic probe with disinfectant coupling agent was carried out sampling survey in a large tertiary firstclass hospital in Hubei Province, bacteria from contaminated probe were counted and identified, SPSS 19.0 software was used for statistical analysis.ResultsA total of 95 B ultrasonic probe specimens were collected, 28.42% were isolated bacteria, the qualified rate was 77.89%; of different departments, qualified rate of B ultrasonic probe in department of gynecology and obstetrics was the highest (92.00%), in physical examination center was the lowest (38.46%); of different types of B ultrasonic probe, qualified rate of transvaginal B ultrasonic probe after covering condom/sterile glove, heart B ultrasonic probe, abdominal B ultrasonic probe, and  transvaginal B ultrasonic probe before covering condom/sterile glove were all≥80%，qualified rate of transvaginal B ultrasonic probe after covering condom/sterile glove was the highest(85.71%），highfrequency probe was the lowest（57.14%），there was no significant difference in qualified rate of different types of B ultrasonic probe (χ2=6.91, P=0.141); 4 strains of methicillinresistant Staphylococcus aureus(MRSA) were isolated, carbapenemresistant Acinetobacter baumannii(CRAB) strain was not found.ConclusionMicrobial contamination exists on B ultrasonic probe with disinfectant coupling agent, health care workers’ awareness on disinfection and isolation should be improved, corresponding cleaning and disinfection system should be set up, monitoring and management should be carried out regularly.