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. 1987 Sep;84(17):6272–6276. doi: 10.1073/pnas.84.17.6272

In situ detection of enteroviral genomes in myocardial cells by nucleic acid hybridization: an approach to the diagnosis of viral heart disease.

R Kandolf, D Ameis, P Kirschner, A Canu, P H Hofschneider
PMCID: PMC299053  PMID: 2819870

Abstract

We have developed an in situ hybridization assay capable of detecting enteroviral RNA in myocardial cells, using molecularly cloned coxsackievirus B3 cDNA as a diagnostic probe. Because of the high degree of nucleic acid sequence homology among the numerous enteroviral serotypes, including the group A and B coxsackieviruses and the echoviruses, detection of these various agents commonly implicated in human viral heart disease is possible in a single hybridization assay. We demonstrate the considerable potential of this method for an unequivocal diagnosis of enteroviral heart disease as well as for pathogenicity studies. Using athymic mice persistently infected with coxsackievirus B3 as a model system, we show that the myocardium is affected in a disseminated, multifocal manner.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abelmann W. H. Viral myocarditis and its sequelae. Annu Rev Med. 1973;24:145–152. doi: 10.1146/annurev.me.24.020173.001045. [DOI] [PubMed] [Google Scholar]
  2. Brahic M., Haase A. T. Detection of viral sequences of low reiteration frequency by in situ hybridization. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6125–6129. doi: 10.1073/pnas.75.12.6125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dec G. W., Jr, Palacios I. F., Fallon J. T., Aretz H. T., Mills J., Lee D. C., Johnson R. A. Active myocarditis in the spectrum of acute dilated cardiomyopathies. Clinical features, histologic correlates, and clinical outcome. N Engl J Med. 1985 Apr 4;312(14):885–890. doi: 10.1056/NEJM198504043121404. [DOI] [PubMed] [Google Scholar]
  4. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  5. Gall J. G., Pardue M. L. Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc Natl Acad Sci U S A. 1969 Jun;63(2):378–383. doi: 10.1073/pnas.63.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hyypiä T., Stålhandske P., Vainionpä R., Pettersson U. Detection of enteroviruses by spot hybridization. J Clin Microbiol. 1984 Mar;19(3):436–438. doi: 10.1128/jcm.19.3.436-438.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Iizuka N., Kuge S., Nomoto A. Complete nucleotide sequence of the genome of coxsackievirus B1. Virology. 1987 Jan;156(1):64–73. doi: 10.1016/0042-6822(87)90436-3. [DOI] [PubMed] [Google Scholar]
  8. Johnson R. A., Palacios I. Dilated cardiomyopathies of the adult (second of two parts). N Engl J Med. 1982 Oct 28;307(18):1119–1126. doi: 10.1056/NEJM198210283071804. [DOI] [PubMed] [Google Scholar]
  9. Kandolf R., Canu A., Hofschneider P. H. Coxsackie B3 virus can replicate in cultured human foetal heart cells and is inhibited by interferon. J Mol Cell Cardiol. 1985 Feb;17(2):167–181. doi: 10.1016/s0022-2828(85)80019-5. [DOI] [PubMed] [Google Scholar]
  10. Kandolf R., Hofschneider P. H. Molecular cloning of the genome of a cardiotropic Coxsackie B3 virus: full-length reverse-transcribed recombinant cDNA generates infectious virus in mammalian cells. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4818–4822. doi: 10.1073/pnas.82.14.4818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kitamura N., Semler B. L., Rothberg P. G., Larsen G. R., Adler C. J., Dorner A. J., Emini E. A., Hanecak R., Lee J. J., van der Werf S. Primary structure, gene organization and polypeptide expression of poliovirus RNA. Nature. 1981 Jun 18;291(5816):547–553. doi: 10.1038/291547a0. [DOI] [PubMed] [Google Scholar]
  12. Lindberg A. M., Stålhandske P. O., Pettersson U. Genome of coxsackievirus B3. Virology. 1987 Jan;156(1):50–63. doi: 10.1016/0042-6822(87)90435-1. [DOI] [PubMed] [Google Scholar]
  13. Nelson J. A., Fleckenstein B., Galloway D. A., McDougall J. K. Transformation of NIH 3T3 cells with cloned fragments of human cytomegalovirus strain AD169. J Virol. 1982 Jul;43(1):83–91. doi: 10.1128/jvi.43.1.83-91.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Racaniello V. R., Baltimore D. Molecular cloning of poliovirus cDNA and determination of the complete nucleotide sequence of the viral genome. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4887–4891. doi: 10.1073/pnas.78.8.4887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Reyes M. P., Lerner A. M. Coxsackievirus myocarditis--with special reference to acute and chronic effects. Prog Cardiovasc Dis. 1985 May-Jun;27(6):373–394. doi: 10.1016/0033-0620(85)90001-5. [DOI] [PubMed] [Google Scholar]
  16. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  17. Rotbart H. A., Levin M. J., Villarreal L. P., Tracy S. M., Semler B. L., Wimmer E. Factors affecting the detection of enteroviruses in cerebrospinal fluid with coxsackievirus B3 and poliovirus 1 cDNA probes. J Clin Microbiol. 1985 Aug;22(2):220–224. doi: 10.1128/jcm.22.2.220-224.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rueckert R. R., Wimmer E. Systematic nomenclature of picornavirus proteins. J Virol. 1984 Jun;50(3):957–959. doi: 10.1128/jvi.50.3.957-959.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tracy S. A comparison of genomic homologies among the coxsackievirus B group: use of fragments of the cloned coxsackievirus B3 genome as probes. J Gen Virol. 1984 Dec;65(Pt 12):2167–2172. doi: 10.1099/0022-1317-65-12-2167. [DOI] [PubMed] [Google Scholar]
  20. Tracy S., Liu H. L., Chapman N. M. Coxsackievirus B3: primary structure of the 5' non-coding and capsid protein-coding regions of the genome. Virus Res. 1985 Oct;3(3):263–270. doi: 10.1016/0168-1702(85)90050-4. [DOI] [PubMed] [Google Scholar]
  21. Wolf H., zur Hausen H., Becker V. EB viral genomes in epithelial nasopharyngeal carcinoma cells. Nat New Biol. 1973 Aug 22;244(138):245–247. doi: 10.1038/newbio244245a0. [DOI] [PubMed] [Google Scholar]

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