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Background  Health workers (HWs) providing routine immunization (RI) services play a crucial role in influencing vaccine uptake, a key determinant of improved immunization coverage. Over the years, Training Needs Assessments (TNAs) have not been routinely utilized in Nigeria to determine unmet needs of health workers offering immunization services and what approaches should be adopted to meet their training needs. The objective was to assess the level of Expanded Program on Immunization (EPI) knowledge among RI service providers and tutors in pre-service institutions in three Nigerian states, to identify unfulfilled training needs and their implications. It also sought HWs perception on a pilot training approach, where tutors will be used for in-service training.    Methods  TNA survey tools were designed to obtain knowledge-based information on the fundamental EPI concepts through key informant interviews and focus group discussions with 90 HWs and 27 pre-service tutors. Quantitative data was also obtained, hence utilizing a mixed method approach for the study.    Results  In spite of several previous trainings, HWs knowledge on basic immunization concepts including Reaching Every Ward (REW) strategy was varied and suboptimal. 83% of the HWs could not differentiate between the live attenuated and killed vaccines. In addition, pre-service tutors knowledge of fundamental EPI concepts, as well as HW perception of the new training approach also varied across the states.    Conclusion  TNAs are valuable in determining specific training approaches to improve HWs skills needed to implement strategies required to increase vaccine uptake. However, EPI managers must be mindful of contextual factors beyond training needs such as finance and security, that can affect HW performance.    Electronic supplementary material  The online version of this article (10.1186/s12913-019-4514-2) contains supplementary material, which is available to authorized users.

Lessons from a training needs assessment to strengthen the capacity of routine immunization service providers in Nigeria