The investigations of ST-segment changes by Holter monitoring demonstrate asymptomatic and symptomatic episodes of myocardial ischemia, which may occur during daily activities. One factor, which is of great importance for the detection of silent myocardial ischemia during ambulatory monitoring, is the combination of the leads. Former studies showed that the analysis of two channels alone may not adequately detect silent myocardial ischemia. We therefore used a three-channel ambulatory ECG monitoring system with a new lead combination. The Holter monitoring results were correlated with the distribution of coronary stenosis detected by coronary angiography. In 54 patients with single coronary vessel disease and ischemic ST-segment depressions during exercise testing, standard Holter lead combination CM2/CM5 was extended by a bipolar Nehb D-like lead. Lead combination CM2/CM5 identified 23 patients (43%) with ST-segment depressions (total number of ischemic episodes = 372). Additional Nehb D-like lead identified 30 patients (55%) with ST-segment depressions (total number of ischemic episodes = 1048). The combination of leads CM2/CM5 and Nehb D raised the number of patients with documented ST-segment depressions to 33 of 54 (61%). Lead Nehb D showed the highest sensitivity for the detection of inferior wall ischemia (stenosis of the right coronary artery); nevertheless, this lead may not be regarded as specific for ST-segment alterations only caused by inferior wall ischemia. The correlation of ischemic ST-segment depressions during exercise testing (classified as anterior, inferior, or anterior and inferior type of ischemia) and documented ST-segment changes in the different Holter leads underline these results. A control group of 40 healthy volunteers demonstrated the high specifity of this new lead combination. In comparison with the standard lead combination CM2/CM5 for the detection of ST-segment changes, lead combination CM2/CM5 extended by a bipolar Nehb D lead is more sensitive for the detection of ST-segment alterations by Holter monitoring.