We examined myocardial tissues for the presence of enteroviral RNA in animal models with experimental coxsackievirus B3 myocarditis and in endomyocardial biopsy samples obtained from patients clinically diagnosed as having dilated cardiomyopathy or myocarditis using polymerase chain reaction (PCR) gene amplification with enterovirus-generic primers and/or coxsackievirus B3-specific primers. In animal models, coxsackievirus B3 was detected in myocardial tissues up to 28 days, 56 days and 180 days after inoculation, in C3H/He mice, A/J mice and Syrian golden hamsters, respectively. The viral genomes were identified by in situ hybridization in myocardial cells and some interstitial cells in and around the myocarditic lesions in animals. In human endomyocardial biopsy samples, enteroviral RNA sequences were detected in 8 (32%) out of 25 patients with clinical dilated cardiomyopathy and in 3 (33%) out of 9 patients with clinical myocarditis. The patients showing histologic findings of myocarditis and clinical features resembling dilated cardiomyopathy had a high incidence (83%) of positive PCR result for enteroviral RNA sequences. Additionally, 25% of patients with dilated cardiomyopathy showing no histologic findings of myocarditis had positive PCR result. This study supports a link between viral myocarditis and dilated cardiomyopathy.