Brucellosis remains one of the major zoonotic diseases worldwide and requires a One Health approach for early detection and control. One of the crucial components for brucellosis control and spread is timely and reliable laboratory diagnosis. External quality assessment is a key component of laboratory quality assurance to evaluate performance and identify possible insufficiencies in laboratory practices. Implementation of brucellosis external quality assessment in resource-limited countries are rare and challenging due to logistical and financial difficulties. The aim of this study was to evaluate the stability of dried tube specimens for external quality assessment of brucellosis serological testing that could be used in resource-limited countries to avoid logistical and financial constraints associated with use of sera. Prepared dried tube specimen panels consisted of 5 samples, one negative and 4 positive samples ranging from weak positive to strong positive. It was shown that brucellosis dried tube specimens were stable at room temperature for 105 days (15 weeks). Consistent results were observed for all samples by rose bengal test during weekly stability testing and also at the end of the stability period by complement fixation test, enzyme-linked immunosorbent assays, and fluorescence polarization assay. In conclusion, brucellosis dried tube specimen maintains integrity of serum samples for serological testing of brucella infection and can be a powerful tool for external quality assessment providers, as it decreases huge shipping costs and avoids challenges in maintaining cold chain shipments between the provider and the recipient laboratories. Moreover, it has great prospects for enabling expansion of external quality assessment programs to include lower tier labs in resource-limited countries to monitor and improve the quality and accuracy of brucellosis testing, as well as it could be used for transportation of clinical samples from remote areas without cold chain logistics.