The present invention relates to a composition used for treating picornavirus infection comprising at least one of the following bacterial strains: Lactobacillus paracasei GMNL-33 with the deposition numbers of CCTCC M 206133, Lactobacillus reuteri GMNL-89 with the deposition numbers of CCTCC M 207154 and Lactobacillus casei GMNL-277 with the deposition numbers of CCTCC M 2013197, and a pharmaceutically acceptable vehicle. The Lactobacillus casei GMNL-277 of the invention is a novel Lactobacillus isolated strain. In addition, the present invention also features the novel use of the composition of the Lactobacillus strains for treating picornavirus infection, and the mechanism of which is inhibition of virus infection by binding of the probiotic bacteria to viruses.