This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kyu, B. Articles by Tracy, S. Search for Related Content PubMed PubMed Citation Articles by Kyu, B. Articles by Tracy, S.

Circulation, Vol 86, 522-530, Copyright © 1992 by American Heart Association

ARTICLES

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

B Kyu, A Matsumori, Y Sato, I Okada, NM Chapman and S Tracy
Department of Internal Medicine, Faculty of Medicine, Kyoto University, Japan.

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 10(2)-10(3) 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.

This article has been cited by other articles:

				H. Higuchi, M. Hara, K. Yamamoto, T. Miyamoto, M. Kinoshita, T. Yamada, K. Uchiyama, and A. Matsumori Mast Cells Play a Critical Role in the Pathogenesis of Viral Myocarditis Circulation, July 22, 2008; 118(4): 363 - 372. [Abstract] [Full Text] [PDF]

				S. Rutschow, J. Li, H.-P. Schultheiss, and M. Pauschinger Myocardial proteases and matrix remodeling in inflammatory heart disease Cardiovasc Res, February 15, 2006; 69(3): 646 - 656. [Abstract] [Full Text] [PDF]

				F. Calabrese and G. Thiene Myocarditis and inflammatory cardiomyopathy: microbiological and molecular biological aspects Cardiovasc Res, October 15, 2003; 60(1): 11 - 25. [Abstract] [Full Text] [PDF]

				L. A. Brewer, H. C. M. Lwamba, M. P. Murtaugh, A. C. Palmenberg, C. Brown, and M. K. Njenga Porcine Encephalomyocarditis Virus Persists in Pig Myocardium and Infects Human Myocardial Cells J. Virol., December 1, 2001; 75(23): 11621 - 11629. [Abstract] [Full Text] [PDF]

				N G Mahon, B Zal, G Arno, P Risley, J Pinto-Basto, W J McKenna, M J Davies, and C Baboonian Absence of viral nucleic acids in early and late dilated cardiomyopathy Heart, December 1, 2001; 86(6): 687 - 692. [Abstract] [Full Text] [PDF]

				K. N. Reetoo, S. A. Osman, S. J. Illavia, C. L. Cameron-Wilson, J. E. Banatvala, and P. Muir Quantitative analysis of viral RNA kinetics in coxsackievirus B3-induced murine myocarditis: biphasic pattern of clearance following acute infection, with persistence of residual viral RNA throughout and beyond the inflammatory phase of disease J. Gen. Virol., November 1, 2000; 81(11): 2755 - 2762. [Abstract] [Full Text]

				C. Kawai From Myocarditis to Cardiomyopathy: Mechanisms of Inflammation and Cell Death : Learning From the Past for the Future Circulation, March 2, 1999; 99(8): 1091 - 1100. [Abstract] [Full Text] [PDF]

				K. O. Schowengerdt, J. Ni, S. W. Denfield, R. J. Gajarski, N. E. Bowles, G. Rosenthal, D. L. Kearney, J. K. Price, B. B. Rogers, G. M. Schauer, et al. Association of Parvovirus B19 Genome in Children With Myocarditis and Cardiac Allograft Rejection : Diagnosis Using the Polymerase Chain Reaction Circulation, November 18, 1997; 96(10): 3549 - 3554. [Abstract] [Full Text]

				T. Shioi, A. Matsumori, and S. Sasayama Persistent Expression of Cytokine in the Chronic Stage of Viral Myocarditis in Mice Circulation, December 1, 1996; 94(11): 2930 - 2937. [Abstract] [Full Text]

				J.-F. Wang, J. Zhang, J.-Y. Min, M. F. Sullivan, C. S. Crumpacker, W. H. Abelmann, and J. P. Morgan Cocaine enhances myocarditis induced by encephalomyocarditis virus in murine model Am J Physiol Heart Circ Physiol, March 1, 2002; 282(3): H956 - H963. [Abstract] [Full Text] [PDF]