['Microvolt T-wave alternans (MTWA) is a valuable tool for stratification of patients at risk for sudden death and has recently been approved for this purpose by Medicare. Although right atrial (RA) pacing has been applied for MTWA testing, the effects of other pacing modalities on MTWA have not been systematically studied. Accordingly, it is unknown whether biventricular (BiV) pacing might influence MTWA test results.', 'This study sought to investigate effects of BiV pacing in comparison with other pacing modalities.', 'Congestive heart failure patients (n = 30) receiving cardiac resynchronization therapy were included, and a systematic step-up pacing protocol was performed via the implanted cardioverter-defibrillator.', 'Of the overall 120 MTWA tests performed, 67 (56%) were nonnegative. Nonnegative MTWA test results were observed in 18 patients (60%) during RA stimulation, whereas 17 (57%), 15 (50%), and 17 test results (57%) were nonnegative during right ventricular (RV), left ventricular (LV), and BiV pacing, respectively. Seven (23%) patients were MTWA negative for all pacing sites. Results of MTWA assessment during RA pacing were concordant with results obtained with RV pacing in 25 (83%) patients (kappa = 0.66, P = .0003), to LV pacing in 21 (70%) patients (kappa = 0.4, P = .025), and to BiV pacing in 25 (83%) patients (kappa = 0.66, P = .0003). Positive and negative predictive values of nonnegative MTWA test results obtained during RA pacing for a similar result obtained with RV pacing were 88% and 76%. Respective values were similar for other pacing modalities (80% and 60% for LV; 88% and 76% for BiV pacing).'] Does biventricular pacing affect microvolt T-wave alternans in heart failure patients?