Background: Healthcare worker (HCW) hands becoming contaminated with microorganisms while providing patient care is thought to be the primary way nosocomial infections spread. Cross-contamination accounts for a large percentage of illnesses, and the primary means of transmission of germs is through the hands of healthcare workers (HCWs). Objective: Isolating and identifying bacterial contamination in hospital staff hands during patient treatment in Shendi hospitals was the aim of this investigation. Methodology: HCWs provided 60 samples between July and September of 2021. Seven different kinds of bacteria were separated from the 59 (98%) positive specimens throughout this investigation. Results: Bacillus cereus 1(1%), Micrococcus luteus 1(1%), Bacillus species 5(7%), coagulase-negative staphylococci 34(48%), staphylococcus aureus 27(38%), Klebsiella pneumoniae 2(3%), and other gram-negative oxidase positive bacilli (unknown) 1(1%) were the most frequently isolated bacterial contaminants from hospital staff hands, according to the results. Antimicrobial susceptibility tests revealed that some of the bacteria recovered from hospital staff member hands are extremely sensitive to imipenem, ciprofloxacin, and gentamicin, respectively, and resistant to ceftriaxone. Conclusion: According to this study, hospital employees at Shendi hospitals may have had bacterial contamination on their hands, which could have resulted in HAIs. Poor hand hygiene habits or the use of non-medicated soap when washing hands could be the cause of the contamination.