We describe 2 patients with endocarditis for whom blood cultures and cardiac valve cultures were repeatedly sterile. Broad-range eubacterial polymerase chain reaction (PCR) amplification performed on cardiac valve specimens from these 2 patients detected DNA of Mycoplasma hominis, for one patient, and of Ureaplasma parvum, for the other patient. Three other cases of infective endocarditis caused by mycoplasmas were identified in the literature. It is important to rule out a diagnosis of mycoplasma endocarditis because the evolution of the disease may be fatal and it requires an adequate and specific antibiotic therapy.