Fire blight, caused by the bacterium Erwinia amylovora, is among the three most important diseases of apple. A major effective method for its integrated management is the reduction of the susceptibility of the host. Cider apple production in Spain is based on local apple cultivars with minimum crop management and phytosanitary control. After the entry of fire blight in Spain, the selection and planting of cultivars with low susceptibility to this disease has thus become of paramount importance. In consequence, and as part of a wider characterization effort, we undertook the evaluation of an apple germplasm collection of local apple cultivars from Spain for susceptibility to fire blight. Because of the quarantine status of E. amylovora in Europe, we evaluated the use of a detached leaf inoculation assay in combination with a traditional shoot inoculation assay to reduce the amount of plant material to evaluate and to minimize pathogen manipulation. Comparison of the susceptibility values for 78 apple cultivars indicated a low but significant correlation (r = 0.56; α = 0.01) between the leaf and shoot inoculation methods. Although the detached leaf assay was not reliable for the direct selection of cultivars with low susceptibility, it was useful to optimize resources and limit the potential dispersal of the pathogen by allowing the exclusion of medium and highly susceptible cultivars from further evaluation. Shoot inoculation of 103 apple cultivars allowed the identification of 48 cultivars with high levels of resistance to fire blight, which could serve as starting material both for apple production and for breeding programs.