Abstract Background Influenza pandemic preparedness plans should be adopted, regularly updated in every country, and comply with international guidelines to ensure proper management of public health emergencies. In Italy, each region develops its pandemic plan based on a National Plan. This study, conducted within a project funded by the Italian Ministry of Health, aims to assess the level of compliance of these plans with the national guidelines. Methods The assessment was based on a checklist of 192 items reporting the actions prescribed by the national plan. The items were divided into macro-areas by pandemic phase: general, alert, inter-pandemic, pandemic, and transition. The compliance regarding the presence and contextualization of each item in the regional plans was evaluated. Each macro-area was further assessed for easy identification of actions, congruence with the National Plan, and contextualization in the regional healthcare system. Some analyses grouped the regions into North, Centre, and South/Islands. Results Some macro-areas, such as ‘Communication’ and ‘Prevention and Control measures,’ showed some weaknesses in all phases, especially in the northern regions, which reached levels of full compliance below 50%. Central regions were highly compliant in almost all macro-areas, especially in ‘Strategy/Governance’ (100%), ‘Supplying’ (75-100%), and ‘Surveillance’ (75-100%). Overall, contextualization was the weakest aspect in all phases, while easy identification and congruence were mainly better achieved. Only three regions fully met all the assessed parameters. Conclusions A strict surveillance and monitoring process of planned activities is necessary to strengthen the country’s preparedness. This study identified critical issues and key areas for improvement at the regional level. This might allow us to verify the correct implementation of national guidelines and define the best strategies for improving readiness for public health emergencies in the future. Key messages • Implementation of the Italian influenza pandemic plan was uneven across regions. • Identification of lacking areas of the regional implementation is essential to improving national preparedness.