Cardiac persistence of cardioviral RNA detected by polymerase chain reaction in a murine model of dilated cardiomyopathy

Bun Sho Kyu, Akira Matsumori, Yukihito Sato, Ikutaro Okada, Nora M. Chapman, Steven Tracy

Research output: Contribution to journalArticle

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Abstract

Background. In our model of dilated cardiomyopathy (DCM), cardiac dilatation and hypertrophy developed after inoculation of encephalomyocarditis virus (EMCV), but the infectious virus was isolated only early after infection. In this study, we investigated whether viral RNA could be detected at later times using the polymerase chain reaction (PCR). Methods and Results. In the in vitro study, FL (human amnion) cells infected with EMCV were harvested for RNA extraction, and viral cDNA was synthesized by reverse transcription with random hexamers. Using oligonucleotide primers with homology to the 5′ noncoding region of EMCV, we enzymatically amplified a 121-base pair band, which was homologous to a probe specific for EMCV as demonstrated by Southern blot hybridization. The sensitivity of this PCR technique was at the level of about 102-103 copies of viral RNA genome. In the in vivo study, four-week-old DBA/2 mice were inoculated with EMCV intraperitoneally (10 pfu/mouse) and killed on days 1, 2, 3, 5, 7, 10, 14, 18, 28, 60, and 90. The hearts were divided into three parts for purification of total RNA, histopathological examination, and to culture for infectious virus. The infectious virus was isolated from the heart after the second day but never after the 14th day. The viral genome was detectable by PCR on the second day, when very little mononuclear cell infiltration around the blood vessels was histologically visible. Positive PCR signals were observed in all hearts through day 14. Viral RNA was also detected in four of six 28-day samples, four of six 60-day samples, and two of seven 90-day samples when diffuse myocardial fibrosis was prominent, but myocardial necrosis or cellular infiltration had disappeared. Conclusions. The persistence of EMCV RNA was shown by PCR in the chronic stage of EMCV-induced myocarditis, a time when the inflammatory reaction had largely subsided. The PCR is a potentially useful method to test possible viral etiologies in idiopathic heart muscle disease or DCM.

Original languageEnglish (US)
Pages (from-to)522-530
Number of pages9
JournalCirculation
Volume86
Issue number2
DOIs
StatePublished - Aug 1992

Fingerprint

Encephalomyocarditis virus
Dilated Cardiomyopathy
DNA-Directed RNA Polymerases
Polymerase Chain Reaction
Viral RNA
Viral Genome
Viruses
RNA
Inbred DBA Mouse
Amnion
DNA Primers
Myocarditis
Cardiomegaly
Southern Blotting
Base Pairing
Reverse Transcription
Blood Vessels
Dilatation
Heart Diseases
Myocardium

Keywords

  • Cardiomyopathy, dilated
  • Myocarditis, viral
  • Polymerase chain reaction
  • Virus, encephalomyocarditis

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

Cardiac persistence of cardioviral RNA detected by polymerase chain reaction in a murine model of dilated cardiomyopathy. / Kyu, Bun Sho; Matsumori, Akira; Sato, Yukihito; Okada, Ikutaro; Chapman, Nora M.; Tracy, Steven.

In: Circulation, Vol. 86, No. 2, 08.1992, p. 522-530.

Research output: Contribution to journalArticle

Kyu, Bun Sho ; Matsumori, Akira ; Sato, Yukihito ; Okada, Ikutaro ; Chapman, Nora M. ; Tracy, Steven. / Cardiac persistence of cardioviral RNA detected by polymerase chain reaction in a murine model of dilated cardiomyopathy. In: Circulation. 1992 ; Vol. 86, No. 2. pp. 522-530.
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