Our objective was to determine if β1-adrenergic receptor (β1-AR) and β2-AR gene polymorphisms influence heart rate (HR), systolic blood pressure (SBP) and diastolic blood pressure (DBP) response to dobutamine during dobutamine stress echocardiography (DSE). Patients (n = 163) undergoing clinically indicated DSE were enrolled. Dobutamine doses were titrated from 5 to 40 μg kg−1 min−1 at 3 min intervals and HR, SBP and DBP were measured. Genotypes were determined for β1-AR Ser49Gly, β1-AR Arg389-Gly, β2-AR Arg16Gly and β2-AR Gln27Glu polymorphisms by polymerase chain reaction-restriction fragment length polymorphism analysis, pyrosequencing and single primer extension methods. β2-AR Glu27 homozygotes had a greater HR response at the highest dobutamine dose than Gln27 carriers (P = 0.002). β2-AR Gly16 homozygotes had a lower HR response during 5-30 μg kg−1 min−1 of the dobutamine infusion protocol compared to Arg16 carriers (P = 0.03). Differences in SBP by β2-AR codon 16 genotype and DBP by β1-AR codon 389 genotype were found at baseline and were maintained throughout DSE (P = 0.06 and 0.02, respectively). However, the magnitude of SBP and DBP response to dobutamine did not differ significantly by β2-AR codon 16 or β1-AR codon 389 genotypes, respectively. These data suggest that the four selected β1- and β2-AR polymorphisms do not substantially influence the magnitude of hemodynamic response to dobutamine during DSE.