Antimicrobial resistance mediated by extended-spectrum beta-lactamases (ESBL), AmpC beta-lactamase and metallo-beta-lactamase (MBL) producing Acinetobacter species is an emerging problem worldwide. In this cross-sectional study total 341 specimens were collected over a period of one year from January 2017 to January 2018. Specimens were collected from ICU and Surgery unit of Mymensingh Medical College Hospital, Mymensingh, Bangladesh. Specimens were collected from ICU and Surgery Unit of Mymensingh Medical College Hospital, Mymensingh, Bangladesh. Samples were processed for culture by standard conventional methods and susceptibility testing and determined by Kirby Bauer disc diffusion method. Antibiotic discs and their strength were according to the CLSI 2017 guideline. Molecular study was done to detect the species by OXA-51 gene and drug resistance genes (IMP, VIM, NDM, TEM, SHV, CTX, SPM, SIM and GIM). Species identification was done by OXA-51 gene which is intrinsic to Acinetobacter baumannii. Among the 46 isolates, 36(78.26%) were positive for Oxa-51 gene, 16(34.8%) for TEM gene, 9(19.6%) for VIM gene, 3(6.5%) for NDM gene and 1(2.2%) for IMP gene. This study gives an alarming sign towards high prevalence of cephalosporin and carbapenem resistance due to production of extended spectrum beta-lactamases and metallo-betalactamases, respectively. Early detection, proper antibiotic policies, and compliance towards infection control practices are the best defenses against these organisms.