Left ventricular (LV) diastolic dysfunction with preserved ejection fraction is frequently encountered in clinical practice. However, the relations between LV filling patterns and myocardial function by using two-dimensional (2D) speckle tracking imaging are not well known. One hundred thirteen individuals (mean age 59 ± 12 years), referred for a clinically indicated echocardiogram, underwent standard and 2D strain echocardiography. The subjects were divided into groups on the basis of diastolic function by using standard Doppler indices, and we compared 2D strain/strain rate parameters between groups. Subjects with mild to moderate LV diastolic dysfunction showed a high left atrial volume index (LAVI) and early mitral inflow to annular velocity ratio. With regard to 2D strain/strain rate, the severity of diastolic dysfunction was associated with low peak strain, and low systolic and early diastolic strain rate of longitudinal, circumferential and radial deformations, whereas the late diastolic strain rate was not. In multivariate analysis, LAVI and circumferential early diastolic strain rate were most associated with the severity of diastolic dysfunction, although longitudinal and circumferential peak strains were also of significant correlation. In addition to LAVI, the circumferential early diastolic strain rate would be a useful parameter in the assessment of the severity of diastolic dysfunction with preserved ejection fraction.