This investigation was to confirm the relationship between asthma, respiratory syncytial virus (RSV) infection, and the gut environment by analyzing the alterations in the gut microbiota of RSV-infected asthmatic mice. All BALB/c male mice were randomly separated into the control group (CON), ovalbumin (OVA) group, and OVA + RSV group (n = 8). At the end of experiment, we evaluated the RSV-infected asthma model using Wright-Giemsa staining, immunoglobulin E (IgE) levels using ELISA, and inflammatory cells using Hematoxylin and eosin (HE). Furthermore, airway hyperresponsiveness (AHR) was measured by a Buxco’s modular and invasive system. Histopathological analysis of the murine lung tissue revealed that the inflammatory infiltration, edema, and collagen hyperplasia were more severe in the OVA + RSV group than in the other groups. Higher expression of interleukin-5 (IL-5), IL-13, IL-25, and IL-33 in mice from the OVA and OVA + RSV groups (*P < 0.05 and **P < 0.01). Moreover, feces were collected for 16S rRNA sequencing and data analysis. The associations between Prevotellaceae_UCG_001, which is positive, and immunoglobulin E (IgE), IL-13, and IL-33 were highly significant (***P < 0.001), IL-5 (**P < 0.01) and IL-25 (*P < 0.05). Lachnospiraceae_NK4A136_group was also positive, and significantly associated with IgE and IL-33. Helicobacter and Uncultured_Bacteroidales_bacterium are negative, had associated with IL-25 (*P < 0.05). Thus, we conclude that asthma with RSV infection can alter the major components of gut microbiota and influence the mutual relationship between the core operational taxonomic units (OTUs) and IgE as well as inflammatory cytokines.