The association of Chlamydia pneumoniae with atherosclerosis is still controversial. Reports from different laboratories have varied widely and "gold standards" for the detection of C. pneumoniae are lacking. In the present study, aortic valves and peripheral blood mononuclear cells from 48 patients undergoing aortic valve replacement were examined for the presence of C. pneumoniae using a nested PCR. C. pneumoniae-specific DNA was not detected in any of the clinical samples. No PCR inhibition was observed by spiking the samples with target C. pneumoniae. A total of 31/46 patients (67%) were seropositive for C. pneumoniae IgG. These results do not support the association of C. pneumoniae with aortic valves and peripheral blood mononuclear cells in patients with atherosclerotic aortic heart valve disease.