Therefore, the findings of this study will highlight the prevalence and associated factors of VF with implications to improve health workers’ interventions, to ensure treatment cost-effectiveness, and to accelerate the reduction of HIV related morbidity and mortality of children. Current and up-to-date information regarding virological failure (VF) in HIV-positive children using routine viral load testing to measure treatment failure is essential for policy makers to take appropriate actions. Though, different studies have been conducted on survival, ART adherence, and other aspects of HIV - infected children, the magnitude and associated factors of VF among children on ART has not yet been well investigated in Ethiopia. Despite hospitals and health centers are using the same ART guidelines, this study included health centers as they cover most health facilities that have been giving pediatric ART services. Furthermore, the previous studies were done at the referral hospitals with diverse health workers to manage cases. However, some critical factors such as HIV status disclosure, level of Hgb, history of ART interruption, CD4 cell counts, and caregiver’s HIV status, which contribute to VF occurrence, were not included. In Ethiopia, few studies have been conducted on TF by considering previous guidelines. Ending the AIDS epidemic by 2030 is the aim of this strategic plan in line with the three 90’s targets. Ethiopia has adopted the UNAIDS 90–90-90 HIV treatment target and has developed HIV/AIDS prevention and strategic treatment plan, which has been implemented since 2015. Likewise, the Ethiopian government launched different strategies, programs, and policies that provide aspiring and far-reaching goals on prevention, treatment, health coverage, and access to affordable medicine for HIV/AIDS patients like Sustainable Development Goals (SDGs). Adherence counseling, early detection of TF, and appropriate switching to second-line therapy are key strengths of a viral load monitoring. In 2014, the Joint United Nations Program on HIV/AIDS (UNAIDS) and partners launched the three 90–90–90 targets, having 90% of patients on ART with VLS is the third “90” targets. The World Health Organization (WHO) recommends routine annual viral load monitoring for all patients on ART, as the most accurate and preferred method to measure treatment response. Treatment failure can be assessed in three ways: clinically, immunologically, and virologically, which offers early and accurate indication of treatment failure. Thus, monitoring people on ART is important to ensure treatment success, identify adherence problems, and determine whether ART regimens should be switched in case of treatment failure (TF). The goal of antiretroviral therapy (ART) is to suppress viral replication and reduce HIV associated mortality among infected children. Beside that 20,130 were newly infected and 7684 were died with HIV related illness. Based on Ethiopian public health institution HIV related estimates and projections about 704,454 populations were living with HIV in 2020. Sub-Saharan Africa (SSA) suffers from the global burden of HIV- infection, with nearly 70% of the world’s HIV/AIDS-infections and deaths occurs in this region. Approximately 1.8 million children (age < 15 years) were living with HIV globally in 2018. Globally, human immunodeficiency virus (HIV) is a major public health problem and associated with a range of short-and long-term consequences. The Creative Commons Public Domain Dedication waiver ( ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made.