Encephalomyocarditis virus RNA was detectable by PCR in murine hearts up to 90 days post-infection, persisting into the chronic stage of myocarditis when infectious virus could no longer be isolated.
Can viral RNA be detected by PCR in the chronic stage of EMCV-induced myocarditis in a murine model?
PCR can detect persistent viral RNA in the chronic stage of viral myocarditis even after infectious virus is no longer isolatable and inflammation has subsided, suggesting its utility in investigating viral etiologies of dilated cardiomyopathy.
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.
Kyu et al. (Sat,) conducted a other in Dilated cardiomyopathy / viral myocarditis. Encephalomyocarditis virus (EMCV) inoculation was evaluated on Detection of viral RNA by PCR at various time points. Encephalomyocarditis virus RNA was detectable by PCR in murine hearts up to 90 days post-infection, persisting into the chronic stage of myocarditis when infectious virus could no longer be isolated.