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Background  Hand hygiene (HH) is recognized as the single most effective strategy for preventing health care–associated infections. In developing countries, data on hand hygiene compliance is available only for few health-care facilities. This study aimed to assess hand hygiene compliance among health-care workers in Debre Berhan referral hospital, Ethiopia.    Methods  This study employed the WHO hand hygiene observation method. Direct observation of the health care workers (HCWs) was conducted using an observation record form in five different wards. Trained and validated observers watched HCWs while they had direct contact with patients or their surroundings, and the observers then recorded all possible hand hygiene opportunities and hand hygiene actions. Observation was conducted over a 24 h period to minimize selection bias. More than 200 opportunities per ward were observed according to WHO recommendation, except in neonatal intensive care unit. HH compliance was calculated by dividing the number of times hand hygiene was performed by the total number of opportunities for hand hygiene. A 95% confidence interval (CI) was computed for compliance with the exact binomial method.    Results  A total of 917 hand hygiene opportunities were observed during the study. Overall HH compliance was 22.0% (95% CI: 19.4–24.9). HH compliance was similar across all professional categories and did not vary by shift. Levels of compliance were lower before patient contact (2.4%; 95% CI: 0.9–5.3), before an aseptic procedure (3.6%; 95% CI: 1.6–7.6) and after contact with patient surroundings (3.3%; 95% CI: 1.2–7.9), whereas better levels of compliance were found after body fluid exposure (75.8%; 95% CI: 68.0–82.3) and after patient contact (42.8%; 95% CI: 35.2–50.7).    Conclusion  HH compliance of HCWs was found to be low in Debre Berhan referral hospital. Compliance with indications that protect patients from infection was lower than that protect the HCWs. The findings of this study indicate that HH compliance needs further improvement.    Electronic supplementary material  The online version of this article (doi:10.1186/s13756-017-0268-y) contains supplementary material, which is available to authorized users.

A twenty-four-hour observational study of hand hygiene compliance among health-care workers in Debre Berhan referral hospital, Ethiopia