Using Balb/c mice infected with Coxsackie virus B3 (CVB3) as a model, the effect of Astragalus membranaceus (AM) on CVB3-RNA has been observed in myocardial tissues of mice by RNA-RNA in situ hybridization with negative-strand RNA probes leballing with 35S and quantitative imaging analysis of positive hybridization signals. The mechanism of its effect on CVB3-RNA has also been investigated by induction with AM and detection of beta-interferon (beta-IFN). Results showed that the copy numbers of CVB3-RNA as well as the histologic necrotic sizes in myocardial tissues of AM treated infected mice were significantly smaller than that in infected normal saline treated mice (P < 0.01, P < 0.05) respectively, suggesting that AM could inhibit the replication of CVB3-RNA, but its effect on CVB3-RNA was not correlated with induction of beta-IFN.