Characterization of coxsackie B virus RNA in myocardium from patients with dilated cardiomyopathy by nucleotide sequencing of reverse transcription-nested polymerase chain reaction products

doi:10.1016/S0046-8177(98)80006-3

Human Pathology

Volume 29, Issue 6, June 1998, Pages 578-584

https://doi.org/10.1016/S0046-8177(98)80006-3 Get rights and content

This study was performed to detect and characterize the enterovirus present in myocardium of some patients with heart muscle disease by nucleotide sequencing of polymerase chain reaction (PCR) products after amplification with enterovirus group-specific primers. Enterovirus sequences have been detected previously in myocardium of patients with myocarditis or dilated cardiomyopathy and seem causal, although the particular virus serotypes involved have not been identified. In a prospective study of endomyocardial biopsy specimens from 35 consecutive patients with suspected heart muscle disease, enterovirus sequences from the 5' nontranslated region were amplified by reverse transcription-nested PCR using group-specific primers. This region contains both conserved and variable sequence motifs, characteristic of particular enterovirus serotypes. The nucleotide sequences of individual PCR products were determined by cycle sequencing and compared with all known sequences (GenBank/EMBOL), using the GCG software package. Endomyocardial biopsy specimens from 9 of 21 (42.9%) patients with a histologically confirmed diagnosis of dilated cardiomyopathy were positive for enterovirus by PCR, compared with only 1 of 14 (7.1%) patients with other myocardial pathological conditions (Fisher's exact probability = 0.0275: odds ratio = 9.75; 95% confidence interval = 1.31−70.78). The nucleotide sequence of the PCR products differed, indicating no cross-contamination. However, computerized comparison showed that each had greatest homology with the 5' nontranslated region of Coxsackie B virus but contained up to 11% sequence variations compared with the prototype Coxsackie B3 strain Nancy. Parallel investigation of tissue from our mouse model of Coxsackievirus B3-induced myocarditis showed that nucleotide sequence changes are not introduced by reverse transcription or PCR. These data support the link between enteroviral infection and dilated heart muscle disease and suggest that Coxsackie B serotypes are the enteroviruses most frequently involved.

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^*: Supported by the British Heart Foundation.

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Original ContributionCharacterization of coxsackie B virus RNA in myocardium from patients with dilated cardiomyopathy by nucleotide sequencing of reverse transcription-nested polymerase chain reaction products*

Lancet

Am Heart J

Lancet

Lancet

Am Heart J

Anal Biochem

Am J Cardiol

J Mol Biol

A six year study of Coxsackie virus B infections in heart disease

J Hyg (Cambs)

In situ detection of enteroviral genomes in myocardial cells by nucleic and hybridisation: An approach to the diagnosis of viral heart disease

Molecular approaches to enteroviral diagnosis in idiopathic dilated cardiomyopathy and myocarditis

J Am Coll Cardiol

Detection of enterovirus RNA in myocardial biopsies from patients with myocarditis and cardiomyopathy using gene amplification by polymerase chain reaction

Circulation

Routine use of muantibody-capture ELISA for the serological diagnosis of Coxsa ckie B virus infections

J Med Virol

End-stage dilated cardiomyopathy: Persistence of enterovirus RNA in the myocardium at cardiac transplantation and lack of immune response

Circulation

Persistance of enterovirus RNA in chronic fatigue syndrome is associated with the abnormal production of equal amounts of positive and negative strands of enteroviral RNA

J Gen Virol

Strand-specific detection of enteroviral RNA in myocardial tissue by in-situ hybridization

Mol Cell Probes

Molecular probes for the detection of persisting enterovirus infection of human heart and their prognostic value

Eur Heart J

Long-term follow-up of acute myocarditis: Correlation of ventricular function and outcome

Eur Heart J

The clinical and prognostic significance of the detection of enterovirus RNA within the myocardium of patients with myocarditis or dilated cardiomyopathy

Circulation

Antibodies to Coxsackie B viruses in primary congestive cardiomyopathy

Br Heart J

Molecular detection and identification of enteroviruses using enzymatic amplification and nucleic acid hybridisation

J Clin Microbiol

Genomic detection of enteroviruses in the myocardium: Studies on animal hearts with Coxsackievirus B3 myocarditis and endomyocardial biopsies from patients with myocarditis and dilated cardiomyopathy

Jpn Circ J

Nested polymerase chain reaction for high-sensitivity detection of enteroviral RNA in biological samples

J Clin Microbiol

Nested PCR for specific detection and rapid identification of human picornaviruses

J Clin Microbiol

Polymerase chain reaction assay for hepatitis C virus RNA using a single tube for reverse transcription and serial rounds of amplification with nested primer pairs

J Med Virol

Original Contribution
Characterization of coxsackie B virus RNA in myocardium from patients with dilated cardiomyopathy by nucleotide sequencing of reverse transcription-nested polymerase chain reaction products*