Background: Plasma is the gold standard specimen forHIV-1 viral load (VL) test.Dried blood spot (DBS) was also recommendedas an alternative specimen by previous studies for this test in resource-limited areas where using plasma is challenging.However, in Ethiopia, where only 77% to 82% the eligible individuals received VL testing, itis not utilizedwell and its performance for HIV-1 VL test was not evaluated. Objective: This study aimed to evaluate the analytical performance of DBSfor HIV -1 viral load test using the Abbott m2000 real-time PCR assay in Bihar Dar, North West Ethiopia. Methods:An institution-based method comparison study was conducted among clients visiting the ART clinics in Bahir Dar from 17 March to 25 September 2025. Finger-prick whole bloodon DBSand plasma in EDTA were collected in parallel from each study participant and analyzed using Abbott m2000 PCR assay protocols for HIV-1 viral load.Testingwas repeated on both samples after a month of storage to assess the effect of testing delay on the results.IBM SPSS version 27 was used for data entry and analysis.Statistical significance was set at p-value<0.05. Results: A total of 189 paired plasma and DBS samples were used in this study.The medians of the differences betweenlog10-transformed DBS and plasma HIV-1 VL resultwas0.013log copies/ml foradjusted data. Chon’s kappa (k) value,for result agreement was 0.87 at 1000copies/ml. We also found, the sensitivity (94.9%); specificity (92.7%); PPV (90.4%) and NPV of (96.2%) at 1000copies/ml,the treatment failure threshold.Similarly,the areas under the receiver operator characteristic (ROC) curve was(0.964) at 1000copies/ml. There was no significant difference between first plasma and repeated DBSVL results after one month storage at ambient temperature, at 1000copies/ml with p=0.727, at α=0.05. Conclusion:The first and after 1month repeated DBS VL results were comparable with first HIV-1 plasma results at 1000 copies/ml. Therefore, DBS can be potentially considered as an alternative option for HIV-1VL test.