Background: The majority of nosocomial infections are seen in intensive care units and they course with higher rates of mortality and morbidity worldwide. Objectives: To identify the current microbial isolates and their antimicrobial susceptibility pattern from the tracheal aspirate culture in a tertiary care hospital in Dhaka city.Materials and Method: This retrospective study was conducted in the department of Microbiology at Holy Family Red Crescent Medical College Hospital, Dhaka from January 2020 to June 2021 for a period of one and a half years. Written consent was taken from the corresponding authority.A total of 109 samples were collected from tracheal aspirates of the patients who were admitted to the hospital with the critically ill patients in intensive care units.Microsoft Excel software was used for data analysis.Results: A total of 109 samples were analyzed. Of them, the predominant populations were male 73(66.97%) and the remaining were female 36(33.03%). Out of 109 samples, 70 (64.22%)was culture positive. Culture negative was 39 (35.78%). The majority of isolates were Gram-negative bacteria. Among them, predominant bacteria wereKlebsiella spp. 43(61.43%) followed by Acinetobacter spp 19(27.15%), Pseudomonas spp. 05(07.14%) & Gram-positive isolates waswereaph. Aureus 03(04.28%). Klebsiella species showed higher sensitivity totigecycline 97.67%, colistin 93.02%, amikacin 65.11% and meropenem 37.20% & gentamicin 30.23%. Other drugs showed wer sensitivity to axicillin/clavulenic acid 18.60%, and ciprofloxacin 18.60% , trimethoprim/ sulfamethoxazole 09.30%, ceftazidime & cefuroxime 04.65% and lowest sensitivity shown in ceftriaxone 02.32%. Acinetobacter speciesshowed higher sensitivityin colistin at 100% and tigecycline at 47.36%. Other drugs like amikacinand trimethoprim/ sulfamethoxazole 15.78% and the lowest sensitivity were shown in meropenem 10.52% and high resistance in gentamycin, ceftriaxone, ceftazidime, and ciprofloxacin.Pseudomonas spp. showed the highest sensitivity 60.00% to piperacillin-tazobactam, amikacin, imipenem, meropenem, and gentamycin, 40.00% to ciprofloxacin, 20.00% to ceftazidime & cefepime.All the isolate of Staph. aureus was the highest sensitive 100% to vancomycin and linezolid, 66.66% sensitive to amikacin, trimethoprim/ sulfamethoxazole, tetracycline, and gentamicin, 33.33% showed lower sensitivityto amoxicillin/ clavulanic acid, cloxacillin and ciprofloxacin.Conclusion:This study aimed to investigate thedistribution of pathogenic microorganisms isolated from tracheal aspirate and their antibiotic sensitivity profile in intensive care units. A periodic review of the bacteriological profile and antibiotic sensitivity pattern is highly essential for the clinician to treat critically ill patients in ICU. Antibiotic policy & infection control programs should be included in every hospital to reduce this drug resistance.