Detection of enteroviruses ribonucleic acid sequences in endomyocardial tissue from adult patients with chronic dilated cardiomyopathy by a rapid RT-PCR and hybridization assay

A rapid reverse transcription polymerase chain reaction (RT-PCR) and microw ell capture hybridisation assay with general specificity for enteroviruses was developed and compared with an improved nested RT-PCR for the detection of enteroviral RNA sequences in endomyocardial tissue from patients with c hronic dilated cardiomyopathy. This method could detect as few as 20 genomi c RNA copies per 100 mg of heart tissue homogenate and results could be obt ained within 8 hours. Of the 55 biopsy specimens aseptically collected from the explanted hearts of 55 patients, 21 (38.2%) were positive by RT-PCR mi croplate assay, whereas only 19 (34.5%) were positive by nested RT-PCR assa y and none were positive by classical cell culture assays. No enterovirus w as detectable by RT-PCR or classical cell culture assays in any of the 55 h eart biopsy specimens taken from organ donors without any known heart disea se. Moreover, the nucleotide sequences of EV nested RT-PCR products showed greatest similarity to group B Coxsackieviruses [CVB3 (n = 12) or CVB5 (n = 3)], but also to group A Coxsackieviruses (CVA21 (n = 1) or CVA9 (n = 3)]. The described RT-PCR and microwell capture hybridisation assay ca n be app lied to the virological diagnosis of human enteroviral cardiac infections. Moreover our findings suggest that group B and group A Coxsackieviruses can persist in heart tissue from patients with endstage chronic cardiomyopathy , supporting the hypothesis that these viruses could be implicated in the e tiology of idiopathic dilated cardiomyopathy. (C) 2001 Wiley-Liss, Inc.