Coxsackieviruses B (CV‑B) are known as the most common viral cause of human heart infections. The aim of the present study was to assess the potential role of CV‑B in the etiology of infectious heart disease in hospitalized patients. The present study is based on blood, pericardial fluid and heart biopsies from 102 patients and 100 control subjects. All of the samples were examined for the detection of specific enteroviral genome using the reverse transcription polymerase chain reaction (RT‑PCR) and sequence analysis. Immunohistochemical investigations for the detection of the enteroviral capsid protein, VP1, from the biopsies were performed. The samples were cultured on confluent KB monolayer cell line for possible virus isolation. The epidemiological data were also collected. CV‑B was detected in 28 of the 102 patients. The sequence analysis demonstrated that 27 strains were identical to CV‑B3 and only one strain was identical to CV‑B1. Furthermore, VP1 in the heart biopsies was detected in enterovirus‑positive cases, as revealed by RT‑PCR. Pericarditis infection was more frequent than myocarditis (P<0.05) or myopericarditis (P=0.05). The epidemiological data demonstrate that CV‑B heart infections occur mainly during autumn and winter, and young male adults are more susceptible than adolescents or adults (P<0.5). The present findings demonstrate a higher prevalence of viral heart infections, suggesting that CV‑B may significantly contribute to heart infections.