BACKGROUND: The debate on the diagnostic value of high intercostal spaces (ICSs) and of the number of diagnostic leads in the Brugada syndrome (BrS) has been settled by a recent expert consensus statement. OBJECTIVE: We have tested the validity of the new ECG diagnostic criteria using echocardiographic, molecular and clinical evidence in one clinical BrS study population. METHODS: We analyzed 114 BrS patients with a spontaneous or drug-induced type 1 pattern recorded in one or more right precordial leads in 4(th), 3(rd) and 2(nd) ICS. The right ventricular outflow tract (RVOT) was localized by echocardiography. All probands were screened on the SCN5A gene. RESULTS: The percentage of mutation carriers (MCs) and the event rate were similar regardless of the diagnostic ICS (4(th) vs high ICSs: MCs 23% vs 19%; event rate 22% vs 28%) and of the number of diagnostic leads (1 vs ≥ 2: MCs 20% vs 22%; event rate 22% vs 27%).The concordance between RVOT anatomical location and the diagnostic ICSs was 86%. The percentage of diagnostic ECG pattern recorded was significantly increased by the exploration of the ICSs showing RVOT at echocardiography (echo-guided vs conventional approach 100% vs 43% p<0.001). CONCLUSION: The high ICSs are not inferior to the standard 4(th) ICS for the ECG diagnosis of BrS and the interindividual variability depends on anatomical location of the RVOT as assessed by echocardiography. This approach significantly increases diagnostic sensitivity without decreasing specificity and fully supports the recently published new diagnostic criteria.