Brucellosis is a zoonotic disease with a high morbidity in developing countries, but there the optimal treatment is not yet determined. Therefore, the development of a simple and effective treatment is important. The aim of this study was to summarize the available evidences and compare rifampicin with streptomycin in human brucellosis with doxycycline as background regimen. We systematically searched PubMed, EmBase, and the Cochrane Library from their inception up through December 2016. We included studies with a randomized controlled design that evaluated the effect of streptomycin compared with rifampicin in human brucellosis patients who received doxycycline therapy as background regimen. The overall failure and relapse were summarized using random-effects model. Our meta-analysis included 1,383 patients with brucellosis from 14 trials. We found that patients who received rifampicin therapy had a higher risk of overall failure (RR: 2.36; 95% CI: 1.72–3.23; P<0.001) and relapse (RR: 2.74; 95% CI: 1.80–4.19; P<0.001) compared with streptomycin. Results of the sensitivity analysis were consistent with the overall analysis. Subgroup analysis indicated that mean age of the patients and percentage of male participants might influence the treatment effects. Furthermore, no publication bias was detected. The findings of this study indicated that rifampicin therapy significantly increased the risk of overall failure and relapse compared with streptomycin. Hence, it can be recommended to patients with human brucellosis receiving streptomycin therapy.