Background: The hygienic status of food handlers is a key determinant of microbial contamination in ready-to-eat foods. Poor hygiene practices can lead to the transmission of pathogenic microorganisms, posing serious public health risks. This study aims to investigate the association between hygiene compliance and microbial contamination in ready-to-eat foods prepared in Khartoum Locality], Sudan. Methods: A cross-sectional study was conducted among (42) food establishments, assessing hygiene using structured observation checklists and testing food samples for Escherichia coli, total coliforms, and Staphylococcus aureus. Microbial counts were compared with hygiene scores using chi-square tests. Results: Food samples from establishments with poor hygiene scores had significantly higher levels of E. coli and total coliforms (p < 0.05). Staphylococcus aureus was detected in [%] of samples from handlers with inadequate personal hygiene practices. A strong correlation (r = [x], p < 0.01) was found between low hygiene scores and elevated microbial counts. Conclusion: The study confirms that substandard hygiene among food handlers is associated with higher microbial contamination in ready-to-eat foods. Targeted interventions to improve hygiene could substantially reduce contamination risks.