Background: Good glycemic control significantly reduces the incidence of diabetesrelated complications; however, it depends greatly on the frequent and accurate monitoring of blood glucose levels. Hand hygiene may determine the accuracy of capillary blood glucose measurements, which may vary due to differences in hand hygiene practices. This study evaluated the impact of different hand hygiene practices on capillary blood glucose levels in Jeddah, Saudi Arabia. Methods: This was an observational cross-sectional study involving the collection of capillary blood samples for the measurement of blood glucose levels of the family medicine residents at King Saud Bin Abdulaziz University in Jeddah, Saudi Arabia. The samples were collected before and after hand washing or after alcohol swab and hand sanitizer use. Results: Of the 98 residents, 69.4% were women, with a mean age of 28.7 ± 3.2 years and age range of 22-37 years. No significant difference was observed in the capillary blood glucose levels of the residents who did not wash their hands compared to that of the residents who used hand sanitizer (P=0.785) or alcohol swabs (P=0.487). Similarly, the blood glucose levels of capillary blood samples from washed hands did not differ significantly from those of samples from unwashed (P=0.227) or sanitized hands (0.270). A significant difference was observable in the blood glucose levels between the first and second drops from unwashed hands (P=0.006) and washed hands (P=0.013), as well as between washed hands and hands on which alcohol swabs were used (P=0.040). Conclusion: Capillary blood glucose levels obtained after using a hand sanitizer, alcohol swabs, or handwashing did not differ significantly from that of samples obtained from unwashed hands. However, a significant difference between the glucose levels in the samples obtained from the first and second drops from both washed and unwashed hands was observed. This suggests that using the second blood drop provides more accurate results.We recommend that patients perform hand hygiene measures and use the second capillary blood drop to obtain a more accurate assessment of blood glucose level. Additional larger studies are required to confirm these findings.