Myocardial ischemia, consisting in a reduction of blood flow to the heart, may cause sudden cardiac death by myocardial infarction or trigger serious abnormal rhythms. Thus, its timely identification is crucial. The Repeated Structuring and Learning Procedure (RS&LP), an innovative constructive algorithm able to dynamically create neural networks (NN) alternating structuring and learning phases, was previously found potentially useful for myocardial ischemia detection. However, performance of created NN depends on three parameters, the values of which need to be set a priori by the user: maximal number of layers (NL), maximal number of initializations (NI) and maximal number of confirmations (NC). A robustness analysis of RS&LP to varying values of NL, NI and NC is fundamental for clinical applications concerning myocardial ischemia detection but was never performed before; thus, it was the aim the present study. Thirteen serial ECG features were extracted by pairs of ECGs belonging to 84 cases (patients with induced myocardial ischemia) and 398 controls (patients with no myocardial ischemia) and used as inputs to learn (50% of population) and test (50% of population) NNs with varying values of NL (1,2,3,4,10), NI (50,250,500,1000,1500) and NC (2,5,10,20,50). Performance of obtained NNs was compared in terms of area under the curve (AUC) of the receiver operating characteristics. Overall, 13 NNs were considered; 12 (92%) were characterized by AUC≥80% and 4 (31%) by AUC≥85%. Thus, RS&LP proved to be robust when creating NNs for detecting of myocardial ischemia.