Most existing masonry buildings were built without following code regulations for seismic criteria. Hence, their performance compared to the demands of the current codes, for both seismic and static actions, could be very low. In engineering practice, strengthening interventions can be pursued at different levels, based on qualitative assessments of masonry structures. Firstly, the structures are evaluated through in situ inspections. Then, based on the structural system of the building a strengthening design is proposed. The design can be targeted at guaranteeing a box behavior of structures acting through macro-elements, or at strengthening the in-plane capacity of piers and spandrels. However, lower cognitive levels may drive inaccurate impressions about the capacity of structures, leading to unreliable strengthening solutions. While in some cases the confidence factor could underestimate the effects of the reinforcements, in other cases lower knowledge levels could lead to incorrect reinforcements that do not provide helpful solutions. In this paper, the issues concerning the strengthening design of masonry structures are presented and discussed with regard to an existing building. The construction is a 20th century masonry building used as a nursing home for elderly persons. The study follows a knowledge-based procedure where, after achievement of the highest knowledge level and the proposal of compatible and effective strengthening interventions, the analysis of the potential effects of achieving lower knowledge levels is discussed both in terms of vulnerability computations and in terms of strengthening effectiveness. Regarding vulnerability assessment, a lower knowledge level leads to non-conservative results, with an increase in the estimated capacity of around 80%. Critical evaluation of the different configurations for the case study makes it clear that for the considered masonry building the mortar investigation represented the most important parameter to be investigated. The proposal of strengthening interventions before assessment of this parameter may lead to ineffective improvements resulting in index values lower than 0.60. The study represents the opportunity to assess comparisons in the setting of different investigation strategies and their effect on the definition of the strengthening solutions.