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a Molecular Pathology
Section, Division of Biomedical Sciences, Imperial College of Science,
Technology and Medicine, Sir Alexander Fleming Building, London
SW7 2AZ, UK, b Key Laboratory of Viral Heart
Diseases of Ministry of Public Health, Shanghai Institute of
Cardiovascular Diseases, Zhongshan Hospital of Shanghai Medical
University, Shanghai, People's Republic of China, c Chuxiong
Institute of Keshan Disease, Chuxiong, Yunnan, People's Republic of
China
Correspondence to: Dr Zhang h.zhang{at}ic.ac.uk
Accepted 21 February
2000
OBJECTIVE
To verify the aetiological
involvement of enterovirus and identify the viral genomic sequences in
Keshan disease.
DESIGNFormalin fixed, paraffin
embedded myocardial necropsy tissue samples were collected in Keshan
disease endemic regions. Fourteen cases with a histologically confirmed
diagnosis of subacute or chronic Keshan disease were studied. Control
tissue included 10 samples of myocardium from cases of cerebral trauma
and one from accidental acid intoxication. One sample from a case of
enteroviral myocarditis was used as a positive control. The presence of
viral genomic RNA was investigated using an established reverse
transcription nested polymerase chain reaction (PCR) coupled with
direct nucleotide sequencing. Further investigations of PCR positive
samples included in situ antigen detection or hybridisation to confirm
positive results.
RESULTSNine of 14 myocardial samples
from Keshan disease cases and the positive control were positive for
the enteroviral RNA. All the controls were negative. Six of the PCR
positive samples were investigated further by in situ enteroviral
antigen or RNA detection and all were positive. DNA sequencing of six
representative PCR products confirmed that they were homologous to the
5' non-translated region of enteroviral genomic RNA. Five had highest
homology to coxsackievirus B genotypes and one was identical to
poliovirus type 3.
CONCLUSIONSThese results support an
aetiological role for enteroviral infection in Keshan disease.
Nucleotide sequence data suggest that coxsackievirus B or coxsackie B
like viruses are often involved in Keshan disease.
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