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Abstract Background Antimicrobial resistance (AMR) is a major public health problem affecting both current and future generations. Development of resistance to the commonly available antibiotics has been directly linked to their irrational use. The World Health Organization (WHO) has clearly stated on the Global Strategy on AMR, judicious antibiotic use as well as adequate knowledge and awareness about AMR, as important tools in curbing the existing problem. Therefore, this study aimed to explore the level of knowledge among accredited drug dispensing outlet (ADDO) dispensers on factors contributing to irrational use of antibiotics among Tanzanian pediatric patients. Methods A qualitative study employing focused group discussions was conducted among 6–10 ADDO dispensers with at least 6 months dispensing experience in 14 regions between July and August 2020. Thematic analysis was used to analyze the data. Results Findings from this study indicate that the ADDO dispensers have adequate knowledge on antimicrobial resistance. Participants were aware of the general meaning and the use of antimicrobials. They were aware that antimicrobials are used for treating infections caused by microbes such as bacteria, fungi, and viruses. Participants knew AMR increases health related costs and that the increasing burden of AMR is not an outcome of a specific group of people such as health care providers or patients, rather a contribution of many factors that bring change in microorganism behavior. Despite this knowledge, participants felt that most of the private health care facilities are profit-oriented; they practice polypharmacy to maximize profit rather than focusing on providing quality health services. Participants revealed that dose shortage after symptomatic relief strongly contributes to AMR. Lastly, the ADDO dispensers expressed that continuous medical education and community education to patients is necessary in the efforts to fight against AMR. Conclusions To mitigate the increasing AMR burden in our society, integrated interventions must include both communities, ADDOs as first point of contact and other healthcare providers. Periodic refresher training with an emphasis on proper practices is crucial to help the dispensers transform their knowledge into action. Responsible authorities should ensure that ADDOs and other private health care facilities adhere to regulations.