Hospital-Acquired Infection (HAI) is one of the highest causes of morbidity and mortality in patients treated in hospitals from bacteria in hospital environments, such as air, water, surfaces, or even medical equipment. The provision of broad-spectrum antibiotics and their combination can cause the bacteria to resist antibiotics. Microbial and antibiotic sensitivity profiling can reduce the resistance rate of bacteria. This research is expected to improve the health status of Indonesian citizens, especially in the area of dr. Soebandi Regional Hospital of Jember. This research used a descriptive research method with a prospective approach using primary data taken from patient's laboratory examinations on aspiration/pus/blood/urine/ sputum/vaginal secretions/CSF/infectious wound/ulcer patients of dr. Soebandi Regional Hospital from January 1st until December 31st, 2019. The patient’s specimen results culture showed positive results in 219 patients. The most common type of infection was sputum infection (43%) and pus infection (42%). The majority type of bacteria is gram-negative (78.5%) and dominated by K. pneumoniae, Escherichia coli, and Pseudomonas aeruginosa. Gram-positive bacteria are found in 47 patients (21.4%), with the most common species are Kocuria varians, Streptococcus sp., and Staphylococcus aureus. The antibiotics with the highest sensitivity were doxycycline for gram-positive and antibiotic for gram-negative are amikacin and piperacillin-tazobactam.