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Background: Despite the existence of national tuberculosis guidelines (NTG) in Ethiopia, the incidence and prevalence of tuberculosis did not decline markedly. Audits could attempt to determine whether or not healthcare professionals actually implemented these guidelines, as non-implementation could contribute to suboptimal tuberculosis treatment outcomes.Aim: To evaluate healthcare providers’ implementation of Ethiopia’s NTG during the diagnosis and treatment of tuberculosis in order to enhance tuberculosis treatment outcomes.Methods: A descriptive, cross-sectional study design was used.Results: Healthcare providers implemented the NTG during tuberculosis diagnosis for female (60.9%; n = 67) and male (56.1%; n = 69) patients. The correct numbers of anti-tuberculosis pills, complying with the NTG recommendations, were prescribed for 91.8% (n = 101) of the women and for 90.2% (n = 111) of the men. However, both over- and under-prescriptions of anti-tuberculosis drugs occurred. There was an over-diagnosis of smear-negative pulmonary tuberculosis. Only 2.6% (n = 2) of the 76 smear-negative pulmonary tuberculosis patients had been diagnosed correctly.Conclusion: Implementation of the NTG should be enhanced, especially with regard to the diagnosis of smear-negative pulmonary tuberculosis patients and the correct prescription of anti-tuberculosis drugs. This would help to increase the number of correctly-diagnosed and -treated tuberculosis patients, improve tuberculosis treatment outcomes, decrease the spread of tuberculosis and prevent the development of multi-drug-resistant tuberculosis strains.